Latest revision as of 04:16, 17 December 2019

Notebook

May

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Wed

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Performers

Philip, Julien

Experiments

gBlocks PCR amplification: IDT-OMPHT,NSP4-tyrosinase,Mefp-5

Protocols

Plasmid Construction

Records

PCR reaction: Prepare reaction mix on ice. Primers for IDT-OMPHT are 100-F/R, for NSP4 tyrosinase are 113/114-F/R, for Mefp-5 are 105-F/R. Concentrations: IDT-OMPHT is 11.0 ng/μl, NSP4-Tyrosinase is 16.7 ng/μl, Mefp-5 is 18.2 ng/μl.

Q5® High-Fidelity 2X Master Mix	25 ul
Primer mix (10cM)	2.5ul
Template (IDT-OMPHT/NSP4 tyrosinase/Mefp-5)	Xμl (We used 10 ng of the fragment as template for each of them)
ddH20	22.5-X
Total	50μl

PCR setting: Tm for IDT-OMPHT is 68°C, for NSP4 tyrosinase is 62°C, for Mefp-5 is 65°C.

Initial denaturation	98°C	30s
X35 Cycles	98°C	10s
	Tm	20s
	72°C	45s
Final extension	72°C	2min
Hold	4°C	Infinite

Results

None

Experiments	052019 gBlocks PCR fragments checking via gel running Repeat gBlocks PCR amplification: IDT-OMPHT,NSP4 tyrosinase,Mefp-5 052191 gBlocks PCR fragments checking via gel running
Protocols	Plasmid Construction
Records	Gel running: Make two different size gels. The small one is 25ml TAE with 0.25g agarose and 1.25ml SYBR safe dye. This gel is for checking the 052019 PCR fragments. The big one is 50ml TAE with 0.5g agarose and 2.5ml SYBR safe dye. This gel is for checking the 052119 PCR fragments. Mix 5μl PCR product and 1μl 6X loading dye. Run at 130V for 30 min and then check the image. PCR reaction and setting: The reaction mix is the same as 052019. Only difference we made is that we changed the Tm into three sets in order to figure out whether Tm led to the incorrect bands for image052019. Tm for Set A is 65°C, Tm for set B is 57°C, and Tm for Set C is 53°C.
Results	After rechecking, we found that we mistakenly ordered the primers for NSP4 tyrosinase. Actually the fragment we ordered has already contained the homologous regions that we wanted.

Experiments

Restriction digestion of plasmid vectors

Recheck gBlocks NSP4 tyrosinase

Protocols

Plasmid Construction

Records

Digestion reaction:

pBAD24 (102.9 ng/μl)	30 μl (30mg of DNA)
EcoRI-HF	1 μl
XbaI	1 μl
Cut smart	5 μl
ddH2O	13 μl
Total	50μl

Incubate at 37°C overnight

Gel running: Run the PCR products of IDT-OMPHT and Mefp-5, and directly run the gBlocks NSP4 tyrosinase at 130V for 30 min and then check the image.

Results

We got the correct band for NSP4 tyrosinase even though it was very weak. We would try to amplify after ligation.

Experiments

Tyrosinase secretion system construction

Protocols

Plasmid Construction

Records

Gel running: 50 μl digested sample + 10 μl loading dye. The order of loading is shown below. Then separate 60μl into two wells on the gel because the well will be overflowing if we put all 60 μl into one well.

1 kb ladder

(2 μl)

Sample

(30 μl)

Sample

(less than 30 μl)

1 kb ladder

(2 μl)

Run at 120V for 23 min in order to separate the digestion products and obtain the target vector (pBAD24, 4516 bp) from unused fragments (26 bp).

Gel purification: We’re using protocol from Illustra GFX PCR DNA and Gel Band Purification Kit Gel. Gel slices weight are 258 mg (used 258 μl CBT3) and 280 mg (used 280 μl CBT3)*CBT3 = capture buffer type 3

Sample capture → sample binding → wash & dry → elution

We used 1 Microspin column and 1 collection tube for purification of 2 gels. Then Elution with 20μl elution buffer type 6

Gibson Assembly reagents calculation:

Insert (NSP4 + tyrosinase)	Vector (pBAD24)
Length: 1004 bp	Length: 4542 bp
Concentration: 16.7 ng/μl	Concentration: 32.80 ng/μl
Volume used for ligation: 5 μl	Volume used for ligation: 3 μl
Mass: 83.5 ng	Mass: 98.4 ng
Mole: 134.6 fmol / 0.1346 pmol (by NEBioCalculator)	Mole: 35.06 fmol / 0.03506 pmol (by NEBioCalculator)

Volume used (Vector : Insert = 1 : 4)

Gibson assembly reaction:

Insert (NSP4 + tyrosinase)	5 μl
Vector (pBAD24)	3 μl
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	2 μl
Total	20 μl

Results

None

Experiments

Tyrosinase secretion system construction

Protocols

Plasmid Construction

Records

Gibson assembly reaction as negative control for 052419:

Master Mix	10 μl
Digested Plasmid	2 μl
ddH2O	18 μl
Total	30 μl

Incubated at 50°C for 15 minutes.

Transformation of NSP4-Tyrosinase-pBAD24 into competent cells:

a. 50 μl competent cells + 5 μl Gibson assembly mix made on 052419

b. 50 μl competent cells + 5 μl negative control made on 052519

c. Spread on two plates with Ampicillin and incubate at 37°C overnight

Results

None

Experiments

Tyrosinase secretion system construction

Protocols

Plasmid Construction

Records

Colony picking and incubation:

Plate	Colony number
Negative control	9 small colonies
pBAD24-NSP4 tyrosinase	1 colony

Prepare the LB Broth with Ampicillin antibiotic and tubes with 1 mL – 5 mL LB Broth with Ampicillin antibiotic.

Pick the only one single colony from the pBAF24-NSP4 tyrosinase transformation plate.

And randomly pick 5 colonies from the negative control transformation plate. Incubate at 37°C overnight.

Results

The negative control has much more colonies than our pBAD24-NSP4 tyrosinase. So we guessed we might mistakenly record the two plates. The results would show us whether our assumption was true after the plasmid extraction and gel running.

Experiments

Tyrosinase secretion system construction

Protocols

Plasmid Construction

Records

Plasmid extraction:

a. Collect 3 mL of overnight suspension culture from the five negative control tubes.

b. Elute with 50μl elution buffer. Did the elution steps three times with the same 50μl elution buffer. Incubate 4min between each centrifuge.

c. Then use Nanodrop to test the extracted plasmid concentration.

Restriction enzyme digestion reaction: Total 50μl

DNA (500 ng)	39.1μl	27.2μl	16μl	18μl	18μl
EcoRI-HF	0.5μl	0.5μl	0.5μl	0.5μl	0.5μl
XbaI	0.5μl	0.5μl	0.5μl	0.5μl	0.5μl
Cut smart	5μl	5μl	5μl	5μl	5μl
ddH2O	4.9μl	16. 8μl	28μl	26μl	26μl

Incubate at 37°C overnight

Results

DNA plasmid extraction: They are 12.81ng/μl, 18.38ng/μl, 31.41ng/μl, 27.2ng/μl and 28.44ng/μl.

Experiments	Tyrosinase secretion system construction
Protocols	Plasmid Construction
Records	Plasmid identity checking for 052619: Run the digested plasmid from five randomly picked colonies on 052619. Loading order for the gel: Left to right 1 kb ladder -> Colony 1 -> Colony 2 -> Colony 3 -> Colony 4 -> Colony 5 Repeat transformation of bacteria: 5 μl Gibson assembly mix made on 052419 and 5 μl negative control made on 052519. Spread on two plates with Ampicillin and incubate at 37°C overnight.
Results	Besides the result of colony 3 that we don’t know how to explain it. Its band is neither the vector nor the NSP4 tyrosinase which was 1022 bp, but this band is around 1500 bp. All the other bands are our vectors which is pBAD24. However, they do not have the insert so they are not the proper plasmid.

Experiments

Tyrosinase secretion system construction

Protocols

Plasmid Construction

Records

Colony picking and incubation

Plate	Colony number
Negative control	3 small colonies
pBAD24-NSP4 tyrosinase	2 colonies

Pick the two colonies from the pBAF24-NSP4 tyrosinase transformation plate.

And pick the 3 colonies from the negative control transformation plate. Incubate at 37°C overnight.

Results

Experiments

Tyrosinase secretion system construction

Protocols

Plasmid Construction

Records

Plasmid extraction:

a. Collect 3 ml of overnight suspension culture from the two pBAD24-NSP4 tyrosinase tubes.

b. Elute with 50μl elution buffer. Did the elution steps three times with the same 50μl elution buffer. Incubate 4min between each centrifuge.

c. Then use Nanodrop to test the extracted plasmid concentration. They are 46ng/μl and 28ng/μl.

Restriction enzyme digestion reaction: Total 25μl

DNA (500 ng)	15μl	18μl
EcoRI-HF	0.5μl	0.5μl
XbaI	0.5μl	0.5μl
Cut smart	2.5μl	2.5μl
ddH2O	10.5μl	3.5μl

Incubate at 37°C overnight

Results

None

June

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Sat

Experiments	Tyrosinase secretion system construction
Protocols	Plasmid Construction
Records	Plasmid identity checking for 053019: Run the digested plasmid from two picked colonies from pBAD24-NSP4 tyrosinase plate on 053019 at 120V for 25 min. Loading order is 1 kb ladder, sample1, sample2, 1 kb ladder. Load 25μl digestion product with 5μl loading dye.
Results	No correct colonies. Through these two transformation experiments, we thought our competent cells have some problem. So we would change to use a new one.

Experiments	Vector identities checking via gel running
Protocols	Plasmid Construction
Records	Expected band size for each plasmid backbone: pBBR1MCS2 = 5148 bp and pBAD24 = 4542 bp I: First gel running around 10 am by Hugh Samples loaded : 10μl pBBR1MCS-2 + 2μl loading dye (x6) 10μl pBAD24 + 2μl loading dye (x6) Ran for 120 V for 25 minutes. Initial observation was that the samples were floating and not settling down even after the addition of the loading dye. Hugh did not run this gel and prepared a new one instead because the TA said maybe the gel preparation is not good. II: Second gel running around 11 am by Hugh Samples loaded: 10μl pBBR1MCS-2 + 2μl loading dye (x6) 10μl pBAD24 + 2μl loading dye (x6) Ran for 120 V for 25 minutes. Gel image is figure 1. III: Third gel running around 5 pm by Philip Samples loaded: 24μl pBBR1MCS-2 + 20μl loading dye (x6) 24μl pBAD24 + 20μl loading dye (x6) Ran for 120 V for 25 minutes. Initial observation: Even after mixing the large volume of loading dye. When loading the sample it was still floating away. We now suspected that there might be something wrong with the sample. We think the sample contains ethanol. Gel image is figure 2. IV: Fourth gel running around 6 pm by Philip Samples loaded: 10μl pBBR1MCS-2 + 2μl loading dye (x6) 10μl pBAD24 + 2μl loading dye (x6) Ran for 120 V for 25 minutes. Initial observation: Same thing as the previous gel No correct band at all, I think it’s due to the loading dye making the fragments too heavy and thus it did not run fast in the gel, therefore our Xiao Xiong (One of our TAs) recommended to prepare a new gel and use a smaller well size. After this result, Xiao Xiong has put the stock plasmid backbone tubes onto a heating block; she opened the cap of the tubes and heated the tubes at 55 C, I reclaimed the stuff after maybe 30 minutes; this was to evaporate possible ethanol contaminants.
Results	Sample might have floated away and none were actually run in the gel. No correct band at all, I think it’s due to the loading dye making the fragments too heavy and thus it did not run fast in the gel, therefore our Xiao Xiong (One of our TAs) recommended to prepare a new gel and use a smaller well size. After this result, Xiao Xiong has put the stock plasmid backbone tubes onto a heating block; she opened the cap of the tubes and heated the tubes at 55 C, I reclaimed the stuff after maybe 30 minutes; this was to evaporate possible ethanol contaminants. It seems that the band is still not at the correct place. There is not even a trace of the correct band for pBAD24 it seems like the band is located more than 10kb. For pBBR1MCS-2, it seems like the DNA sample all floated away and what we saw that settled were probably just the loading dye. Next time we should mix the loading dye and the DNA sample better. Maybe there is something wrong with the plasmid? After consultation with Professor Zheng Jun’s PhD. Student, apparently we should first digest the plasmid vector with at least one enzyme to linearize it. Only then should we run it in the gel and check the size. Apparently if we do not linearize the plasmid before running, due to its circular form it would run slower along the gel causing it to show a non-true band.

Experiments

Repeat vector identities checking via gel running

Protocols

Plasmid Construction

Records

Restriction enzyme digestion reaction for vectors: Total 10μl A1 is pBBR1MCS-2 041719, A2 is pBBR1MCS-2 050519, B1 is pBAD24 041719, B2 is pBAD24 050519.

Vector	A1	A2	A3	A4
DNA (500 ng)	4μl	8.5μl	1μl	5μl
EcoRI-HF	0.5μl	0.5μl	0.5μl	0.5μl
Cut Smart	1μl	1μl	1μl	1μl
ddH20	4.5μl	0μl	7.5μl	3.5μl

Incubate at 37℃ for 1h. Gel running: 120V for 25 min. 10μl digestion product + 2μl loading dye. Gel arrangement is 1kb ladder, A1, B1, B2, A2, 1 kb ladder. Restriction enzyme digestion reaction for insert: Total 50μl. Incubate overnight.

Insert DNA (200 ng)	8.17μl
EcoRI-HF	2μl
Xbal	2ul
ddH20	37.83μl
Cut Smart	5μl

Results

After digestion with EcoRI-HF for 1 hr at 37 C. We ran the plasmid stock that has kept (041719) along with the stock plasmid solution that has prepared (050519). The 041719 are at the expected size: pBBR1MCS2 = 5148 bp and pBAD24 = 4542 bp.

Experiments	Vector and insert checking via gel running
Protocols	Plasmid Construction
Records	Sample:55μl digested insert + 10μl loading dye Order: 1 kb ladder(μl), sample(30μl), sample(30μl), 1 kb ladder(2μl) Run at 120V for 30 min. Our expected size is 4515bp for vector pBAD24 and 1004bp for NSP4+tyrosinase
Results	The left one is for vector pBAD24 and the other one is for insert NSP4+tyrosinase. The right one was the supposedly digested insert but we did not see any band there.

Experiments	pBAD24, pBBR1MCS2 plasmid extraction for sequencing
Protocols	Plasmid Construction
Records	Pellet 4 ml of the 5 ml overnight culture. Pipette 1 ml into microcentrifuge tube and centrifuge for 3 min. Discard supernatant and add next 1 ml into same tube. Repeat until you’ve pelleted 4 ml worth of bacteria culture. (12 min of centrifugation) Follow protocol in QIAGEN (miniprep) plasmid extraction kit. *Miniprep is for 5 ml bacteria culture, yield is low because starting vol. is low. Mediprep is for 50 ml, maxi for 100, and giga for 1L. P1 is resuspension buffer. P2 is lysis buffer. (Alkaline, with many salts) N3 is neutralization buffer. (Balance alkalinity) You’d see some white precipitate after adding N3. The centrifuge step in protocol is to spin down most of that ppt. for you to collect supernatant. This prevents clogging (blocking) of the spin column for downstream procedure. Nanodrop at Prof. Xu’s Lab. Clean with water and blank, then measure. pBAD24: 66.4ng/μl (acceptable) pBBR1MCS2: 37.6ng/μl (still pretty low) Afternoon: Philip sent them for plasmid sequencing.
Results	None

Experiments

Resuspension of gBlock fragments

Protocols

Plasmid Construction

Records

Centrifuged the tube at 4500 x g to make sure the dried pellet is at the bottom of the tube. Added 100μl TE buffer to all of the fragments (I was expecting to get 10ng/μl). All products that were delivered were 1000 ng. Vortexed briefly. Incubated at 50 C for 20 minutes. Vortexed then centrifuged. Got the concentration using Nanodrop, we used TE buffer as blank. After the last step, I aliquoted 50μl to another tube so that we would avoid exposing the main stock to many freeze/thaw cycles which could damage the DNA too. So there is about 50μl fragment volume left on the main IDT tubes that we have received.

Results

Resulting concentration:

Concentration	Aliquoted tube (50μl)	IDT Tube (~50μl)
NSP4-Tyrosinase (1022 bp)	10.84 ng/μl = 0.017 pmol	8.341 ng/μl = 0.013 pmol
OPHT (1495 bp)	8.82 ng/μl = 0.0095 pmol	7.147 ng/μl = 0.0077 pmol
OPHTII (2211 bp)	7.65 ng/μl = 0.0055 pmol	6.968 ng/μl = 0.0075 pmol
OPHTIII (1507 bp)	5.68 ng/μl = 0.0060 pmol	23.29 ng/μl = 0.02501 pmol
OPHM (1782 bp)	3.85 ng/μl = 0.00349 pmol	5.164 ng/μl = 0.0047 pmol

Pmol calculations through https://nebiocalculator.neb.com/#!/dsdnaamt.

Experiments

Restriction digestion of pBBR1MCS2

Protocols

Plasmid Construction

Records

Restriction digestion reaction: (pBBR1MCS2 concentration:75.5ng/μl)

pBBR1MCS2(2.5mg)	2500/75.5≈33ml
EcoRI	3μl
BamHI	3μl
Cut smart	5μl
dd H2O	6μl
Total	50μl

Put in thermocycler for 1h. Settings are as following: 37℃ for 1h, 4℃ for infinite

Gel running: Loading samples (50μl+10 loading dye). Put original plasmid to compare with digested one. Run at 120V for 20 min.

Gel purification: After excise bands, we did gel purification using protocol from Illustra GFX PCR DNA & Gel Band Purification Kit Gel. Final elution volume is 40μl. Then using Nanodrop to check its concentration which is 7.278ng/μl. It is not a good concentration result. So we repeated the experiment again.

Repeat restriction digestion: Prepare 4 tubes, 50μl each, total 200μl. The reaction is the same as previous. Overnight digestion at 37℃.

Results

Both are the digested products, we just separated them into two wells like usual because one well might overflow and we would lose DNA. Both of them pBBR1MCS2 are at the expected size.

Experiments	Repeat gel purification
Protocols	Plasmid Construction
Records	Gel running: This time we put all digested products into one big well. (200μl sample+40μl loading dye). Order is 1kb ladder, sample, and 1 kb ladder. Then Still run at 120V for 20 min. Gel purification: After excising the gel and following the same protocol, we eluted it with 20μl elution buffer.
Results

Experiments

pBBR1MCS2+OPHTIII construction

Protocols

Plasmid Construction

Records

Gibson assembly:

Insert (OPHT III)	Vector (pBBR1MCS2)
Length: 1507 bp	Length: 5148 bp
Concentration: 23.29 ng/μl	Concentration: 52.3 ng/μl
Volume used for ligation: 3.6 μl	Volume used for ligation: 1.91 μl
Mass: 83.5 ng	Mass: 98.4 ng
Mole: 25.01fmol /0.025 pmol (by NEBioCalculator)	Mole: 16.44 fmol / 0.016 pmol (by NEBioCalculator)

Volume used (Vector : Insert = 1 : 3)

Gibson assembly reaction:

Insert (OPHT III)	3.6μl
Vector (pBBR1MCS2)	1.91μl
Gibson Assembly Master Mix (2X)	10μl
Deionized H2O	4.49μl
Total	20 μl

Incubate at 50℃ for 1h.

Transformation of bacteria: The only changes that we usually make are the following:

Incubation times for both thawing and incubating with DNA. Varies from 25-30 mins
The amount of LB broth to put for the outgrowth step varies from 500-900 ul (This case Hugh used 500 ul)
Incubation time for the outgrowth step varies from 45 mins - 1 hr (This time we used 1 hr)
Centrifuge 3000 x g for 2 minutes after outgrowth step to pellet the bacteria then remove some supernatant to leave 250 ul LB solution then resuspend the pellet with the leftover 250 ul LB solution then plate all the bacteria on the plate.
Dry the plates for 10 minutes before inverting and putting into the 37 C incubator

Results

None

Experiments	pBBR1MCS2+OPHTIII construction
Protocols	Plasmid Construction
Records	Colony picking and incubation Picked 6 colonies from the plate Put 8 mL LB broth total and added 8 uL Ampicillin Put in 37 C shaking incubator overnight for 12-16 hrs
Results	None

Experiments	pBBR1MCS2+OPHTIII construction
Protocols	Plasmid Construction
Records	Plasmid extraction: using QIAprep Spin Miniprep Kit. 6 tubes containing 8ml transformed bacteria cultured with LB broth. Pellet 6ml bacterial overnight culture from each tube. Add 250μl buffer D1, then 250μl buffer D2, 350μl buffer N3. Centrifuge at 13000 rpm for 10 min. Transform the 800μl supernatant into spin column. Centrifuge at 13000 rpm for 60s and discard flow through. Add 500μl buffer PB with ethanol. Then centrifuge again at 13000 rpm for 60s and discard flow through. Add 750μl buffer PE, centrifuge at 13000 rpm for 60s and repeat twice. Put the spin columns into new tubes and add 35μl buffer EB. Stand for 5min then centrifuge again. Using Nanodrop to measure their concentration.
Results	None

Experiments	pBBR1MCS2+OPHTII Construction
Protocols	Plasmid Construction
Records	Gibson assembly Transformation of bacteria
Results	Failed: subsequent days were used as diagnostic and review

Experiments	pBBR1MCS2+OPHTII Construction
Protocols	Plasmid Construction
Records	Colony picking and overnight culture
Results	None

Experiments	Repeat pBBR1MCS2+OPHTII Construction
Protocols	Plasmid Construction
Records	Gibson assembly Transformation of bacteria
Results	None

Experiments	Repeat pBBR1MCS2+OPHTII Construction
Protocols	Plasmid Construction
Records	Plasmid Extraction of pBBR1MCS2: using Midi Kit Transfer the overnight culture into four 50ml tubes. Nanodrop result is 54.42 ng/ul Restriction digestion PCR for 2 hours at 37 degrees, infinite hold at 4 degree Gel run for 25 mins at 130V (We didnt save the gel image results), we cut the lower band and did gel purification for the pBBR1MCS2 vector.
Results	None

Experiments	Repeat pBBR1MCS2+OPHTII Construction, Bacteria Transformation
Protocols	Plasmid Construction
Records	Gibson Assembly of pBBR1MCS2 and OPHT2 Run for 1 hour at 50 degrees, infinite hold at 4 degrees Transformation: We thawed the competent cells for only 12 minutes, we should thaw them for a longer time. Then we spread the bacteria on Kan+ agar plates, culture them for 16 hours.
Results	Transformation: It has no colony on positive control plate but few colonies on OPHT2 plate.

July

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Experiments

Repeat pBBR1MCS2+OPHTII Construction, Start OPHM construction and Transformation of OPHM

Protocols

Plasmid Construction

Records

Colony picking and bacteria incubation: Picked 4 colonies from OPHTII agar plate for overnight LB broth culture LB broth volume 40ml + Kanamycin volume 0.04ml

Gibson assembly for pBBR1MCS2+OPHM construction:

(insert) OPHM concentration = 5.164ng/ul = 0.0047pmol

(vector) digested pBBR1MCS2 = 12.60ng/ul = 0.004pmol

60ng of vector = 60/12.60 = 4.76ul

4.76ul x 0.004pmol = 0.019 pmol

vector:insert=1:2

Insert = 0.019pmol x 2 =0.038pmol

Volume for insert = 0.038/0.0047 = 8.09ul

pBBR1MCS2+OPHM (vector:insert=1:2)

Insert (OPHM)	8.09 μl
60ng Vector (pBBR1MCS2)	4.76 μl
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	0 μl
Total	22.85 μl

Positive control

Insert	0 μl
Positive control	10ul
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	0 μl
Total	20 μl

Negative control

Insert	0 μl
Vector (pBBR1MCS2)	10ul
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	0 μl
Total	20 μl

Incubate at 50℃ for 1h.

Transformation:

a. Heat shock at 42℃ for 45 secs using water bath.

b. Incubate at 37℃ for 60 mins, then plated them into three plates and cultured them for 16 hours.

Results

Transformation: The positive control plate has no colony due to our mistake of plating the bacteria to Kan+ agar plate, it should be plated on Amp+ agar plate.The negative control plate has many colonies which should not happen, we suspect that the plasmid is self-ligated although we transformed with vector without insert. The OPHM agar plate has man colonies which is the expected results.

Experiments

Repeat pBBR1MCS2+OPHTII Construction  Plasmid Extraction of pBBR1MCS2-OPHT2

Restriction digestion for plasmid identity checking

Protocols

Plasmid Construction

Records

Plasmid Extraction: We eluted with 40ul elution buffer for x3 times.

Restriction digestion reaction:

	C1	C2	C3	C4
500ng DNA	500/82.28=6.1ul	500/87.78=5.7ul	500/85.39=5.9ul	500/88.74=5.6ul
EcoRI	0.5μl	0.5μl	0.5μl	0.5μl
BamHI	0.5μl	0.5μl	0.5μl	0.5μl
Cut smart	1μl	1μl	1μl	1μl
dd H2O	1.9μl	2.3μl	2.1μl	2.4μl
Total	10μl	10μl	10μl	10μl

Put in thermocycler for 2hrs. Settings are as following: 37℃ for 2hrs, 4℃ for infinite hold.

Gel running: Load 4 samples (10μl+2 loading dye) and 1kb ladder (2ul) into wells. Put original plasmid to compare with digested one. Run at 120V for 20 min.

Restriction digestion reaction: (pBBR1MCS2 concentration:75.5ng/μl)

pBBR1MCS2(2ug)	40ul
EcoRI	1μl
BamHI	1μl
Cut smart	5μl
dd H2O	3μl
Total	50μl

We made two tubes of the reaction above. Put in thermocycler for 14hrs. Settings are as following: 37℃ for 14hrs, 4℃ for infinite.

Results

Concentration of OPHT2 plasmid:

colony 1(C1)=82.28ng/ul

colony 2(C2)=87.78ng/ul

colony 3(C3)=85.39ng/ul

colony 4(C4)=88.74ng/ul

Gel run: The expected results should be two bands shown on the gel as the plasmids should be digested by enzyme at the restriction sites. However, there was only a band for each sample, it might due to the plasmid is undigested or not fully digested. Therefore, we have to reconstruct the plasmid backbone.

Experiments

Repeat pBBR1MCS2+OPHTII Construction

pBBR1MCS2+OPHM Construction

Prepare LB agar plates

Protocols

Plasmid Construction

Records

Gel purification: Run gel. Order is 1kb ladder, pBBR1MCS2, pBBR1MCS2, none, OPHT II, pBBR1MCS2, pBBR1MCS2. Used 20μl elution buffer type 6. Concentration is 25.42ng/μl.

Gibson assembly reaction:

OPHT II concentration: 7.645ng/μl = 0.0055pmol

pBBR1MCS2 concentration: 25.42ng/μl = 0.008pmol

OPHM concentration: 5.164ng/μl = 0.0047pmol

1.pBBR1MCS2+OPHTII (insert: vector=3:1)

Insert (OPHT II)	8.7 μl
100 ng Vector (pBBR1MCS2)	3 μl
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	0 μl
Total	21.7 μl

pBBR1MCS2+OPHM (insert: vector=2:1)

100 ng Insert (OPHM)	10.21 μl
Vector (pBBR1MCS2)	3 μl
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	0 μl
Total	23.2 μl

Negative control

Insert	0 μl
Vector (pBBR1MCS2)	10ul?
Gibson Assembly Master Mix (2X)	10 μl
Deionized H2O	0 μl
Total	20 μl

Incubate at 50℃ for 1h.

Transformation of OPHT2, OPHM and negative control:

a. Heat shock at 42℃ for 45secs using water bath.

b. Incubate at 37℃ for 45 min then plated them into three plates.

c. Culture them for 12 hrs.

Prepare LB agar plates: 200ml LB agar= 8g powder+ 200ml MilliQ water.

The ratio for antibiotics and solution is 1:1000. So 200μl Amp/Kan to each kind of plate.

We made totally 18 plates with Kan+ and 21 plates with Amp+.

Results

None

Experiments

Colony picking for OPHT2, OPHM Transformation of OPHT1, OPHT3 and NSP4-Tyrosinase

Amplify the plasmid

Plasmid Extraction of OPHT2, OPHM

Restriction digestion to check plasmid identity

Protocols

Plasmid Construction

Records

Colony picking: We picked 6 colonies from each transformed plasmids and incubate them in 5mL LB with antibiotic for 12-14 hrs.

Transformation: Transform OPHT2(#2), OPHT3(#2), NSP4(#6) into Dh5 strain E.coli and incubate at 37 degrees.

Amplify the plasmid by streaking bacteria glycerol stock and culture in 5mL LB broth with antibiotic.

Plasmid Extraction: Save overnight culture before extraction. Used 30ul elution buffer and eluted 3 times, each time incubated 3 minutes. Concentration of DNA is measured by using Nanodrop.

Restriction digestion reaction:

	#1	#2	#3	#4	#5	#6
500ng OPHM	500/95.65=5.3ul	5.3ul	5.3ul	4.8ul	5.1ul	5.5ul
EcoRI	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl
BamHI	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl
Cut smart	1μl	1μl	1μl	1μl	1μl	1μl
dd H2O	3.1μl	3.1μl	3.1μl	3.6μl	3.3ul	2.9ul
Total	10μl	10μl	10μl	10μl	10μl	10μl

	#1	#2	#3	#4	#5	#6
500ng OPHT2	4.3ul	4.1ul	5.2ul	4.6ul	4.7ul	4.9ul
EcoRI	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl
BamHI	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl	0.3μl
Cut smart	1μl	1μl	1μl	1μl	1μl	1μl
dd H2O	4.1ul	4.3ul	3.2ul	3.8ul	3.7ul	3.5ul
Total	10μl	10μl	10μl	10μl	10μl	10μl

Put in thermocycler for 4hrs. Settings are as following: 37℃ for 4hrs, 4℃ for infinite hold.

Run the samples on the agarose gel and observe the DNA bands under UV. The estimated correct band size of plasmid colonies are sent for RNA sequencing.

Results

Concentration of DNA:

OPHM OPHT2

#1=95.65ng/ul #1=117.1ng/ul

#2=94.81ng/ul #2=124.3ng/ul

#3=94.47ng/ul #3=97.22ng/ul

#4=204.6ng/ul #4=109.4ng/ul

#5=98.68ng/ul #5=108.2ng/ul

#6=91.51ng/ul #6=102.3ng/ul

Gel run and observation:

OPHT2 (#2 ans #5)

OPHM(#2 and #6)

Performers

Philip, Julien

Experiments

1.Preparation of 20% Arabinose

2.SDS-Dye preparation

3.Protein samples preparations

Protocols

Plasmid Construction

Records

1: Prepared 5 mL 20% arabinose

Mixed 1051.9 mg Arabinose + 5 mL milliQ water

2: 4x SDS sdye preparation

Component name	Volume
4 x stacking gel buffer	25 mL
SDS	2 g
Glycerol	20 mL
Bromophenol blue 2-mercaptoethanol	20 mg

3: Protein samples preparation

OPHT, OPHTIIII and NSP4 Pellets

-Resuspended them all with 75 ul Milli-Q water + 25 ul 4 x SDS Page dye. Stored in -20C fridge

Stored 1 mL of the supernatant of the induced culture tube for later protein expression test.

Prepared loading sample for supernatant. 75 ul NSP4 supernatant + 25 ul 4x SDS page dye

Results

None

Performers	Renee
Experiments	pBBR1MCS2+OPHTII(sequenced) Construction pBBR1MCS2+OPHM(sequenced) Construction
Protocols	Plasmid Construction
Records	Transformation: 2.5ul OPHTII #5 with 50ul competent cell 2ul OPHM #2 with 50ul competent cell Plated at 16:15 and incubated at 37 degree overnight.
Results	None

Performers

Philip

Experiments

SDS-Page gel running and Coomassie blue staining

Protocols

Protein Expression

Records

Gel percentage used 12.5%

Protein samples that were ran:

NSP4 = 37.17 kDA

OPHTI = 52.18 kDA

OPHTIII = 52.83 kDA

10x Prepared running buffer:

Component name	Amount
Tris	30g
Glycine	144g
SDS	10g

Gel running arrangement:

NSP4 supernatant -> NSP4 pellet -> OPHT3 -> OPHT -> Ladder -> NSP4 supernatant -> NSP4 pellet -> OPHT3 -> OPHT -> Ladder

Running time: 60V for 1hr then 120 V for 1 hr

Coomasie blue staining:

Washed the SDS away from the gel
Soaked the gel with the gel stain for 1 hr
Wash the excess gel stain with dH2O.
Put water after 3 times washing, boil for 5 minutes in the microwave then let it shake overnight. Start time 10:27 pm
Remove the water and take picture

Results

None

Performers	Philip, Renee
Experiments	Make glycerol stock of pBBR1MCS2+OPHTII(sequenced) and pBBR1MCS2+OPHM (sequenced)
Protocols	Plasmid Construction
Records	200ul 5% glycerol + 800ul bacteria culture Separate 10 tubes SDS dye
Results	None

Performers	Renee
Experiments	Test adhesion system in three cell lines
Protocols	Plasmid Construction
Records	Transformation: TOP10 OPHT#1(1ul) OPHT3 #3(1ul) 07/05 vector (3.5ul) BL-21 OPHT#1(1ul) OPHT3 #3(1ul) 07/05 vector (3.5ul) MG1655 OPHT#1 (1ul) OPHT3 #3(1ul) 07/05 vector (3.5ul)
Results	None

Performers	Renee
Experiments	Test adhesion system in three cell lines
Protocols	Plasmid Construction
Records	Pick colony: three colonies per plate for TOP10 and BL-21 with OPHT, OPHT3 and vector, totally 18 tubes, culture at 37 degree shaker overnight Transformation: TOP10 OPHT2 OPHM BL-21 OPHT2 OPHM MG1655 OPHT2 OPHM
Results	No colony grew on all MG1655 plates. Repeat the plating with all the remainder MG1655 culture.

Performers	Renee
Experiments	Test adhesion system in three cell lines
Protocols	Plasmid Construction
Records	Pick colony: three colonies per plate for TOP10 and BL-21 with OPHT2 and OPHM, totally 12 tubes, culture at 37 degree shaker overnight
Results	Still no colony grew on all MG1655 plates. MG1655 was not suitable for our expression system.

August

Sun

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Fri

Sat

Performers	Renee, Philip
Experiments	pet11a+OPHT/OPHT2/OPHT3/OPHM/OPHA1/OPHA2 Construction
Protocols	Plasmid Construction
Records	Gel making Gel running: 50ul PCR product + 10ul loading dye, load 30ul per well for pet11a+OPHT/OPHT2/OPHT3/OPHM/OPHA1/OPHA2; 3x50ul PCR product + 30ul loading dye, load 90ul per well for pet11a vector. Run at 120V 15mins Loading order: Gel1-ladder/OPHT1/OPHT1/OPHT1/OPHT2/OPHT2 Gel2-blank/ladder/OPHT3/OPHT3/OPHM/OPHM Gel3-ladder/OPHA1/OPHA1/blank/OPHA2/OPHA2 Gel4-ladder/vector/vector Gel cut weight: OPHT1 0.36g Buffer type 3: 360ul OPHT2 0.27g 270ul OPHT3 0.31g 310ul OPHM 0.31g 310ul OPHA1 0.16g 160ul Vector1 0.57g 570ul Vector2 0.43g 430ul Concentration: OPHT1 41ng/ul OPHT2 38.36ng/ul OPHT3 18.95ng/ul OPHM 21.67ng/ul OPHA1 10.02ng/ul Vector1+ Vector2 54.12ng/ul
Results	No band for OPHA2

Performers

Philip, Julien, Renee

Experiments

Repeat pet11a-OPHA2 Construction

Continue pet11a-OPHT/OPHT2/OPHT3/OPHM/OPHA1 Construction

Protocols

Plasmid Construction

Records

1. OPHA2 PCR followed by running gel

2. Gibson assembly:

OPHT1	1.91ul
pet11a vector (100ng)	1.85ul
H2O	6.24ul

OPHT2	3ul
pet11a vector (100ng)	1.85ul
H2O	5.15ul

OPHT3	4.2ul
pet11a vector (100ng)	1.85ul
H2O	3.95ul

OPHM	4.2ul
pet11a vector (100ng)	1.85ul
H2O	3.95ul

OPHA1	7.63ul
pet11a vector (100ng)	1.85ul
H2O	0.52ul

3. Transformation: 2ul DNA+50ul competent cell Stbl3 Transformed OPHT1, OPHT2, OPHT3, OPHM, OPHA1 and positive ctrl; plated 300ul transformed bacteria on agar plates

Results

Still no band showed on the gel for OPHA2

Performers

Philip, Julien

Experiments

Plasmid Extraction

Protocols

Plasmid Construction

Records

Used 30ul elution buffer for elution

Results

DNA Concentration (ng/ul):

OPHT #1	109.3
OPHT #2	156.8
OPHT #3	116.4
OPHT2 #1	124.6
OPHT2 #2	116.5
OPHT2 #3	147.4
OPHT3 #1	107.9
OPHT3 #2	105.8
OPHT3 #3	140.9
OPHM #1	148.0
OPHM #2	94.47
OPHM #3	118.0
OPHA #1	126.6
OPHA #2	83.39
OPHA #3	114.0

Performers	Philip, Julien
Experiments	Tyrosinase Activity Assay
Protocols	Functional Test
Records	Sample 1: Induced NSP4 pellet Sample 2: Induced NSP4 supernatant Sample 3: Uninduced NSP4 pellet Sample 4: Uninduced NSP4 supernatant Homogenize cells with 500ul ice-cold Tyrosinase Assay buffer to perform lysis and keep on ice for 10 minutes followed by centrifugation at 10000xg for 15 minutes at 4 degree. Collect the supernatant and estimate protein concentration by using BCA protein assay kit (protein concentration should range between 1 to 2.5 ug/ul)
Results	None

September

Sun

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Tue

Wed

Thu

Fri

Sat

Performers	Philip, Julien, Renee
Experiments	Prepare NSP4 for Negative control NSP4 activity test preparation
Protocols	Protein Expression
Records	Took one tip of cells from glycerol stock, incubate with 5ml LB broth + 5ul Ampicillin at 37 degree shaker at 17:05 Pull down assay for NSP4 induced supernatant. Sonication for NSP4 bacteria culture
Results	None

Performers

Philip, Julien

Experiments

Nickel pull-down assay

Tyrosinase activity test

Protocols

Protein Expression

Records

Prepare nickel resin: Pipette 30ul of nickel resin. Wash resin in 1ul H2O two times followed by washing with 1ml binding buffer two times.
Pull-down assay: Centrifuge the induced BL-21 (NSP4 transformed colony 3) as well as the uninduced one and take the supernatant as pull-down assay sample. Pipette 30ul resuspend resin into supernatant. Incubate and rotate it at 4 degree for 45mins. Spin at 3900rpm for 10mins, remove the supernatant which will be stored just in case, then leave 1ml fraction in resuspend resin and transfer resin to 1.5ml tube. Spin down at 4000g 2min to get resin. Discard supernatant. Wash resin with 750ul binding buffer three times (we washed the resin with Tyrosinase Assay buffer in the third wash). Measure protein concentrationProceed to Tyrosinase Activity Assay.
Tyrosinase activity test: Homogenize cells with 500ul ice-cold Tyrosinase Assay buffer to perform lysis and keep on ice for 10 minutes followed by centrifugation at 10000xg for 15 minutes at 4 degree. Collect the supernatant and estimate protein concentration by using BCA protein assay kit (protein concentration should range between 1 to 2.5 ug/ul)

Results

Protein	Concentration
Uninduced pellet	1.67ug/ul
Induced pellet	5.64 ug/ul
Uninduced supernatant	-0.19ug/ul
Induced supernatant	-0.11ug/ul

As the induced supernatant concentration is negative, there is literally no protein in the supernatant, therefore we did not continue proceed the Tyrosinase Activity test.

Performers	Julien
Experiments	Make an overnight culture of BL-21 (NSP4 transformed colony 3) from glycerol stock
Protocols	Plasmid Construction
Records	Make a 20mL culture of BL-21 glycerol stock (NSP4 transformed colony 3) with Ampicillin antibody (1:1000 dilution) and start incubation in 37 degree shaking incubator at 6.25pm
Results	None

Performers	Julien, Renee
Experiments	pBAD24 NSP4 #3 transformation with BL-21 Induce overnight BL-21 (NSP4 transformed colony 3)
Protocols	Plasmid Construction
Records	Transformation: 1.5ul pBAD24 NSP4 #3 + 50ul BL-21 competent cell Induction: separate 5ml out of 20ml overnight BL-21 as uninduced BL-21 stock and put into -80 degree, for the rest of 15ml, add 150ul 20% arabinose for induction, then culture overnight in 37 degree shaker
Results	None

Performers	Philip, Julien, Renee
Experiments	Preparation for NSP4 activity assay Colony picking
Protocols	Plasmid Construction
Records	Picked a colony from the transformation yesterday. Prepared a 20 mL culture with amp+ and incubated in 37 C at 6:49 am Centrifuge the overnight induced culture of NSP4 Colony #3 BL-21, separate pellet and supernatant, then store in -80degree. Centrifuged 5mL uninduced sample NSP4_plasmid#3 then stored in -80C. Induced the rest 15 mL (OD=1.1227 13-14 hr culture) with 150 ul 20% arabinose at 20:08 and incubated in shaking incubator.
Results	None

Performers	Philip, Julien, Renee
Experiments	NSP4 activity assay
Protocols	Protein Expression
Records	Nickel pull down both induced and uninduced Nsp4 SUPERNATANT (new and old stock), total 4 samples Scrapped OPHT2 bacteria (three stocks) from glycerol stock 082719 and cultured in ~5mL volume of Lb broth with amp+ and then put in 37C shaking incubator at 16:45. Glycerol stock for all OPHT2 colonies were thawed completely by accident watch out for the quality of this stock for next scrapping. Homogenize 4 samples with tyrosinase assay buffer and test protein concentration with BCA protein assay (30mins incubation)
Results	None

Performers	Philip
Experiments	OPHT2 induction NSP4 activity assay Material stock preparation
Protocols	Plasmid Construction Functional Test
Records	Induced 5mL cultures of pet11a_OPHT2 from yesterday with 1mM IPTG (10 ul of 500mM IPTG stock) and incubated at 37 C shaking incubator at 4:45. OD600 =0.7144 Tyrosinase activity assay started plate OD monitoring (510nm) at 8:02 am in the plate reader in 37 C temperature for 1 hr. Stored all proteins back in the “Protein sample box” in -80C Made two 500mL LB broth and one 110 mL LB agar for making 10 amp+ plates. Made a stock of 50 mL LB Broth amp+ and put in 4 degree fridge. Diluted induced pet11a_OPHT2 #1 from early this morning. 1:1000 dilution ratio by putting 10 ul of that induced culture into 10 mL LB Broth amp+ then stored in shaking incubator 37 C overnight.
Results	Result of tyrosinase activity test today

Performers	Philip
Experiments	BL-21 OPHT2 #1 EGFP expression
Protocols	Plasmid Construction Functional Test
Records	eGFP expression is still very high even after ~28 hrs after induction for our bacteria Bl-21 OPHT2 colony #1
Results

Performers	Philip, Julien
Experiments	Material stock preparation BL-21 OPHT2 #1 EGFP expression Adhesion system reconstruction
Protocols	Plasmid Construction
Records	Made 11 Amp+ agar plates by adding 100ul (1:1000) to 100ml agar solution. eGFP expression is still very high even after ~40 hrs after induction for our bacteria Bl-21 OPHT2 colony #1 PCR reaction for the amplification of adhesion system for ligation to pet11a for 1 hr and 26 mins. Started at 15:34
Results

Performers

Philip, Julien, Renee, Jia Ying

Experiments

Adhesion system reconstruction

Preparation digested pet11a vector stock

Protocols

Plasmid Construction

Records

1. Incubated a new BL_21 OPHT2 colony #1, 5 mL volume, at 37 C

2. Run the gel with opht1,2,3 and opha1 samples (each well 30ul, each sample 2 wells) at 120V for 20 mins. Then gel purification.

3. Gibson assembly to ligate Pet11a vector with Opht1,2,3 and Opha1 inserts. Calculation as below:

Plasmid	Concentration(ng/ul)	pMol
pet11a vector	54.12	0.015
OPHT1	6.7	0.0068
OPHT2	7.4	0.0051
OPHT3	4.5	0.0045
OPHA1	17.3	0.018

75ng of vector=75/54.12=1.39ul

1.39ulx0.015pmol=0.02pmol

vector:insert=1:3

Insert=0.06pmol

Insert	Volume of insert(ul)	Volume of vector(ul)	Volume of water(ul)	Volume of Master Mix(ul)	Total volume (ul)
OPHT1	8.82	1.39	-	10	20.21
OPHT2	11.76	1.39	-	13	26.15
OPHT3	13.33	1.39	-	14	28.72
OPHA1	3.33	1.39	5.28	10	20

4.Restriction digestion for pet11a vector. We did it for 100ul, incubate for 14hrs at 37 degree, infinite hold at 4 degree. Calculation as below:

pET11a concentration: 49.1ng/ul

pET11a (3ug)	60ul
BamHI	2ul
NHeI	2ul
Cutsmart buffer	10ul
ddH2O	26ul
Total	100ul

5. Transformation stock OPHT1, OPHT2, OPHT3, OPHM, and OPHA using BL-21; stock pBBR1MCS2_OPHT2 using BL-21; stock pet11a_OPHT1, OPHT2 using Rosetta. Put into the incubator at 21:00

Results

Transformation:

Performers	Julien, Renee, Jia Ying
Experiments	Colony picking for transformed bacteria Bacteria transformation Ammonia iron (III) citrate solution preparation Induction of IPTG and Ammonia iron (III) citrate in OPHT2 bacteria Gel run and gel purification
Protocols	Plasmid Construction
Records	Colony picking: Picked 1 colony for OPHT1,2,3,M and OPHA1 (BL-21), OPHT1,2 (Rosetta). Picked 3 colonies for pBBR1-Opht2 (BL-21). Incubate the picked colonies with 5ml Amp+ (for pET11a plasmid) or Kan+ (for pBBR1MCS2 plasmid) LB broth, put in 37degree shaking incubator at 4.30pm. Transformation with new constructed pet11a_OPHT, OPHT2, OPHT3 and OPHA using the Stbl3 strain E.coli. Put them into 37 degree incubator at 18:40. Prepared the Ammonia iron (III) citrate solution at concentration:100mM with 26.3g iron(III) powder in 263mL water at 18:00. We added 10ul IPTG and 1ml ammonium iron(iii) citrate into BL-21 OPHT2 at 6pm. We run the gel for digested pET11a vector, the expected band size is 5.6kb. Then we purify the excised gel (band1 weight 0.371g and band2 weight 0.372g) with 30ul elution buffer type 4 three times, each time incubated 5minutes.
Results	Gel run results: The concentration for band1 is 2.24ng/ul and band 2 is 2.49ng/ul.

Performers	Philip, Julien
Experiments	Glycerol stock making Colony picking and bacteria overnight culture
Protocols	Plasmid Construction
Records	Picked up adhesion system plasmids transformed plates from 37C, parafilmed and put it in 4C. All constructs have colonies. Picked three colonies for each transformed plasmids in Stbl3 with 5ml LB, incubated in 37 shaking incubator at 6.50pm. Collected the overnight culture of picked colonies at 1.30pm [amplified adhesion system-old stock: OPHT1,2,3,M and OPHA1 (BL-21), OPHT1,2 (Rosetta) and pBBR1-Opht2 #1,2,3 (BL-21)]. Made 1ml glycerol stock of these with 500ul 50% glycerol and 500ul bacteria culture, stored in -80 degree fridge’s Glycerol stock box.
Results	Checked EGFP expression of pbbr1-opht2 with fluorescence microscope:

Performers	Julien
Experiments	Sequencing samples preparation Glycerol stock making
Protocols	Plasmid Construction
Records	Collected overnight culture of picked colonies (pet11a-stbl3) and prepared sequencing samples(1ml bacteria, 120ul 106FOR, 30ul 102 FOR), sent for sequencing. Made 1ml glycerol stock with 500ul 50% glycerol stock and 500ul bacteria culture, stored in -80 degree fridge’s glycerol stock box.
Results	None

Performers	Julien
Experiments	Protein extraction
Protocols	Protein Expression
Records	Protein extraction for overnight bacteria culture (pet11a adhesion system-BL21, Rosetta) with 500ul lysis buffer, boiled at 98degree for 10minutes. Measured protein concentration after 30minutes BCA incubation. Run western blot at 1.40pm. The order and volume loaded shown as below: Ladder - OPHT1 (Rosetta) - OPHT2(Rosetta) - OPHT1 (BL-21) - OPHT2 (BL-21) - OPHT3 - OPHM - OPHA1 (5ul for ladder, 20ul for the rest protein samples) Incubated with antibody and shaking in 4 degrees fridge at 5.20pm
Results	None

Performers	Philip, Julien
Experiments	Continue western blot
Protocols	Protein Expression
Records	Washed the membrane with about 3 times. Incubated 10 mL OPHT2 Colony #1 in LB amp+ at 37 C shaking incubator at 23:30. Magnetization of E.coli incubated in Ferric ammonium citrate(3 days incubation at 37 C) for 3 hrs with magnet. Made a video from the fluorescence microscope
Results	Western Blot results:

Performers	Philip
Experiments	Incubation of bacteria culture, adhesion system functional test
Protocols	Functional Test
Records	Incubated 15 mL culture of all the new pET11a adhesion system glycerol stock at 37 C shaking incubator at 14:01. Will extract after 12 hrs. Induced the overnight BL-21 with 1mM IPTG (50ul in 10 mL) and put in 37C shaking incubator. Inducing for 3 hrs. Added 2 ml of 100mM ferric ammonium citrate into the 3 hr IPTG induced BL-21 OPHT2 colony #1(Checked egfp signal and it’s positive) and put it in 37C shaking incubator at 17:08. Incubating for 24 hrs. Made 5 mL overnight culture of BL-21 for adhesion system functional test. Stored in 37 C shaking incubator at 20:50 . Used all colony #1 from the old pet11a glycerol stock. Incubating for 12 hrs.
Results	None

Performers	Philip
Experiments	Plasmid extraction Induction of adhesion system Magnetization test set up
Protocols	Plasmid Construction Functional Test
Records	Extracted plasmids of the 15 mL new pet11a adhesion system and sent samples for sequencing. Stored new pet11a plasmids in -20C in the new construct box. Induced 5 mL overnight for functional test of adhesion system using 1mM IPTG(10 ul 500 mM stock -> 5 mL culture) for 3 hrs in 30C shaking incubator in Prof.Xu’s lab started at 16:30. Started the magnetization test setups at 00:05. Prepared three dishes. Two that was incubated with fe3+ and one negative ctrl. One of the two dishes has been put in the fluorescence microscope and picture is being taken at 30 sec intervals for 6 hrs.
Results	None

Performers	Philip
Experiments	Incubation of bacteria culture
Protocols	Plasmid Construction
Records	Started the incubation of overnight culture of Tyrosinase (500 U/ml) 2 mL + 2 mL of our adhesion system bacteria that was resuspended with 10 uM Phosphate buffer (Which prepared at 092419). Incubated in Professor Xu's lab's shaking incubator temperature 25-27 C, shaking at 75 rpm. Picked up our overnight culture E.coli with Tyrosinase at 19:45 (Total 17 hrs incubation). Started the incubation of our "Activated E.coli with Tyrosinase" at 20:30. Mixed 150 ul of the nanoparticle PFODTBT (30 ppm) with the 4 mL overnight culture. Put the solution in Professor Xu's shaking incubator at 25C 70 rpm for 1 hr. Put the activated bacteria incubated with the nanoparticle in the 4 Degree of PRofessor Xu's lab.
Results

Performers	Julien
Experiments	Checking of nanoparticle binding with bacteria
Protocols	Plasmid Construction
Records	Centrifuged the nanoparticles incubated bacteria culture and resuspended the pellet with 1mL phosphate buffer. Get 10ul on glass slide and observed under a fluorescence microscope. We have negative ctrl(without tyrosinase), nanoparticles only and 5 bacteria samples.
Results	None

Performers	Julien
Experiments	Bacteria culture Restriction digestion
Protocols	Plasmid Construction
Records	Prepare 5mL of pet11a adhesion system BL-21 bacteria culture with Amp+, shaking in 37 incubator of Prof. Henry’s lab. Restriction digestion for pBAD4 vector and pBAD4-tyrosine vector with XbaI enzyme. Put in thermocycler 37 degree for 14 hours and 4 degree for infinite hold.
Results	None

Performers

Philip, Julien, Renee

Experiments

DNA purification

Induction of samples

Gibson assembly

Transformation

Protocols

Plasmid Construction

Records

Separated 1ml culture as uninduced sample for each. Induced with 10uL IPTG for each samples except negative ctrl and incubated in Prof’s Xu lab 30 degree shaking incubator at around 11.40pm.

Purified DNA of digested pBAD24 and pBAD24_NSP4 from agarose gels. Concentration of pBAD24 is 125.1ng/ul and pBAD24_NSP4 is 61.31ng/ul.

Resuspend ftnA fragment with 25ul TE buffer followed by 50degree incubation for 20min. Gibson assembly of ftnA+pBAD24 and ftnA+pBAD24_NSP4.

Gibson assembly for pBAD vector+ftnA and pBAD NSP4 +ftnA. Put in thermocycler for 1hr at 50degree. Calculation as below:

pBAD NSP4 conc=61.31ng/ul = 0.017pmol

pBAD conc=125.1ng/ul = 0.042pmol

ftnA conc=20ng/ul = 0.052pmol

100ng vector (pBAD NSP4)=1.63ul

(vector)1.63 x 0.017=0.028pmol

vector:insert=1:3

(insert)=0.083pmol

ftnA volume=0.083/0.052=1.6ul

Vector	1.63ul
Insert	1.6ul
Master mix	10ul
water	6.77ul
total	20ul

100ng vector(pBAD)=0.8ul

(vector)0.8 x 0.042=0.034pmol

vector:insert=1:3

(insert)=0.102pmol

ftnA volume=0.102/0.052=1.96ul

Vector	0.8ul
Insert	1.96ul
Master mix	10ul
water	7.29ul
total	20ul

Incubated our 1 mL bacteria with tyrosinase 500U/mL in Professor Xu’s shaking incubator at 25 C 90 rpm for 24 hrs.

Transformation of ftnA+pBAD24 and ftnA+pBAD24_NSP4 into stbl3 strain. Start incubation at 22:17.

Results

None

Performers	Philip
Experiments	Colony picking
Protocols	Plasmid Construction
Records	Picked three colonies for FtnA+pBAD24 and made 25 mL bacteria solution with amp+ at 17:30. Unfortunately there were no colonies for the pBAD24_NSP4+FtnA. Will repeat this transformation later.
Results	None

Performers	Philip, Julien
Experiments	Plasmid extraction
Protocols	Plasmid Construction
Records	Plasmid extraction for three overnight culture of pbad24-ftna. Used 30ul EB, repeated 3times and each time incubated 5 mins. Extracted concentration listed in "Results" Incubated 100 ul of activated adhesion system + bacteria from 092819 with 10 ul (1 mg/mL concentration) of 1.0%w/v FP-00552-2 Yellow fluoresscent particle, 0.04-0.09 um size in 25C and mixing at 300 rpm for 1 hr. Started at 23:40 (Samples were covered with aluminum foil to prevent exposure to light).
Results	Concentration of extracted plasmid: pBAD24-ftnA #1: 169.3ng/ul pBAD24-ftnA #2: 200.9ng/ul pBAD24-ftnA #3: 137.8ng/ul

October

Sun

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Sat

Performers	Philip
Experiments	Adhesion functon test
Protocols	Functional Test
Records	Adhesion test fluorescence checking using 4th floor plate reader machine. Result: Failed will elaborate later. Third functional test using 092819 1000ul activated bacteria. Put 5 ul of the yellow nanoparticle from Prof Xuanjun 30ppm into the 1000ul bacteria. Incubated at 25C for 1hr with shaking for 400 rpm. Started at 19:00. Incubated new adhesion system bacteria for functional test. Picked all Colony #1. Prepared 5 mL overnight culture amp+, will subculture tomorrow to 20 mL culture (Need more bacteria concentration for the tests). Started the incubation at 22:30 at shaking incubator 37 C.
Results	None

Performers	Philip
Experiments	Incubation of bacteria culture Induction of adhesion system
Protocols	Plasmid Construction Functional Test
Records	Subcultured 5 mL overnight cultures into 20 mL LB amp+ and incubated at 37 C for 3 hrs. Started incubation at 13:00. Saved the 5 mL overnight culture as uninduced samples. Will measure OD of both later, need OD=2 for activation and functional tests. Induced Subcultured 20 mL adhesion system bacteria culture with 1 mM IPTG each (OPHT, OPHT2, OPHT3, OPHM, & OPHA). Incubated at 30 C for 4 hrs in Professor Xu's shaking incubator. Update on the magnetization effect after incubation with ammonium ferric citrate of the BL-21 strain of E.coli. They can still be attracted to magnet until now since 092419. Update on adhesion 20 mL culture, I will induce it overnight instead of 4 hrs. Will harvest them tomorrow morning instead.
Results	None

Performers	Philip
Experiments	Checked sequences of newly constructed plasmid
Protocols	Plasmid Construction
Records	Checked the sequences of the newly constructed pet11a adhesion system plasmids. Conclusions are as follows： OPHT1 colony # 3 = Should be correct (Has a single point mutation but it's peak looks like a sequencing error) OPHT2 colony #1 = Correct and usable if needed OPHT3 = no correct or intact colony OPHA Colony #1 and #3 = Correct and usable if needed
Results	None

Performers	Philip
Experiments	Sent sample for sequencing
Protocols	Plasmid Construction
Records	Send ftna samples that Julien extracted from 093019 to Tech dragon sequencing.
Results	None

Performers	Philip, Renee
Experiments	Functional test of the combination of tyrosinase and adhesion system
Protocols	Functional Test
Records	Incubated 100ul bacteria with 50ul 1500U/ml tyrosinase and 1ul of 1%w/v yellow nanoparticle together for 4 hrs and 8hrs. Will check the fluorescence reading after these two time periods. Cultured 20 mL bacteria using glycerol stock of colony number 1 for OPHT, OPHT2,OPHT3, OPHM, OPHA1 with Amp+ and one for BL21 only without Amp. Current conclusions for adhesion system assay parameters: bacteria amount (Need OD=2), time of incubation with tyrosinase (4hrs, is the best I think), amount of tyrosinase (U/mL) (500U/ml or less), amount of nanoparticle (10ml/L or 10ppm is good for our experiment), centrifugation speed(3000xg 3 mins no nanoparticle pelleting), learning the plate reader machine(440 nm first filter and 480nm second filter), choosing the nanoparticle(1% w/v Yellow fluorescent particle from spherotech and red nanoparticle of Prof Xuan Jun), assay format in 96 well plate (As shown in the picture just now), Amount of bacteria and nanoparticle total volume (150 ul) so we can plate in 96 well. ligated pBAD24_NSP4 with fntA, and a negative control only has pBAD_NSP4 without insert then transformed them into DH5 alpha. Started incubation at 20:23. Cultured 18 tubes 20ml of bacteria from glycerol stock NSP4-Tyrosinase colony 3.
Results	Current conclusion based on this fluorescence plate reader result is that after 4 hrs of incubation with tyrosinase there seems to be a significant decrease of supernatant fluorescence signal compared to our inactivated supernatant samples. Most notable OPHM.

Performers	Philip, Julien, Renee
Experiments	Induction of NSP4 tyrosinanse
Protocols	Functional Test
Records	Induced with 5 concentrations of arabinose in 15 tubes of nsp4 tyrosinase bacteria overnight culture. Another 3 tubes are not induced with arabinose act as negative ctrl. The volume of arabinose added shown as below: Induced overnight OPHT1, OPHT2, OPHT3, OPHM and OPHA with 1 mM 40 ul of (500mM IPTG) per 20 mL culture and incubated in Professor Xu's shaking incubator 30 C for 4 hrs. Started ~approximately 12:08. Collected 4 hr incubation with 5 concentrations arabinose’s bacteria culture. Nickel pull down for nsp4 4 hr arabinose induced (supernatant only), stored the pulled down protein in -80c protein samples box. Collected 8hr samples and centrifuging at 3000xg for 10 mins. Will separate supernatant into another tube. Incubated adhesion 100219 and 100519 bacteria with yellow nanoparticle started incubation at approx~ 21:00. Checked the construction of three extracted plasmids pbad24-fntA on 093019 by PCR and gel running. Nickel pull down for nsp4 8 hr arabinose induced (supernatant only), stored the pulled down protein in -80 protein samples box. Collected 12hr samples and centrifuging at 3000xg for 10 mins. Stored separated-supernatant into another tube and stored in -80c.
Results	Nanoparticle Fluorescent signal of inactivated and activated supernatant of OPHT, OPHT2, OPHT3, OPHM, OPHA, Negative control, and Positive control.

Performers

Philip

Experiments

Incubation of bacteria containing FtnA plasmid

Functional test of adhesion system

Tyrosinase activity assay

Protocols

Functional Test

Records

Incubated transformed bl-21 with FtnA plasmids. Diluted outgrowth solution 1:100 and plated 350 ul of the bacteria. Put it in Prof Xu’s 37 degree incubator at 1:16 am.

Incubated BL-21 with pBAD24_FtnA at 14:20 in Professor Xu's 37 C incubator. Only plated pBAD24_FtnA#1 and pBAD24_FtnA#2.

Tyrosinase activity assay for 4,8,12 hours incubation with arabinose at 5 different concentrations. We cultured them at 30 degree accidentally.

Tyrosinase activity assay reagents set-up:

Sample Background Control (SBC) mix: add this into SBC wells

Assay buffer	45ul x 18 = 810ul
Enhancer	5ul x 18 = 90ul

Reaction mix: add this into assay background control, samples and positive control wells

Assay buffer	35ul x 21 = 735ul
Substrate	10ul x 21 = 210ul
Enhancer	5ul x 21 = 105ul

Positive control well: 2ul positive control + 48ul assay buffer

Assay background control: 50ul Assay buffer

Plate:

Results

Results for the adhesion system functional test. Conclusion: The results are quite consistent. Plate was read three times with the same machine and averaged all the reads as shown in the picture data set. Then I averaged each the three repeats for each samples and obtained final reads for each 100219 and 100519 samples and made the graph based on them. We can see a clear difference with the activated and inactivated one. Negative control (Wild type, just BL-21) is too close to our inactivated samples, not sure if the activated readings would be considered significant, but this negative control sample has been prepared along with the 100519 sample so it actually cannot act as a negative control for the 100219 samples but I just put it anyway. We need a third biological repeat for all these to have a final conclusion. Anyways, the data is very consistent, activated sample's readings are always lower than the inactivated ones for these first two biological repeats.

The results of plates after 1 hour measurement in the plate reader (30seconds interval, 37 degree incubation, OD 510nm) is saved.

Performers

Julien

Experiments

Nickel pull-down for NSP4 tyrosinase protein

Protocols

Protein Expression

Records

Incubated 15 tubes of 5ml nsp4 bacteria with 5 different arabinose concentration (2%, 0.2%, 0.02%, 0.002% and 0.0002%) at 12.30pm.

Concentrations of arabinose	Volume to add
2%	500ul
0.2%	50ul
0.02%	5ul
0.002%	0.5ul
0.0002%	0.05ul

Collected nsp4 tyrosinase bacteria after 4hr incubation (at 4.30pm), centrifuged and keep the supernatant. Nickel pull down for 4hr incubation batch nsp4 bacteria’s protein and stored in -80 fridge’s bacteria box. (Because no space in protein sample box). Collected 8 hr incubation of different concentrations arabinose in nsp4 at 8.30pm. nickel pull down for 8hr incubation batch nsp4 bacteria’s protein and stored in -80 fridge’s bacteria box. (Cuz no space in protein sample box). Collected 12 hr arabinose incubation at 5 different concentrations of nsp4 bacteria at 12.30am . Centrifuged and kept the supernatant in -80 with a green holder.

Results

None

Performers	Philip, Julien
Experiments	Nickel pull down
Protocols	Protein Expression
Records	Nickel pull down Diluted protein samples by 10x and measure the protein concentration using BCA assay.
Results	Protein concentration:

Performers

Philip, Julien, Pinto

Experiments

Tyrosinase activity assay

Protein extraction

Western Blot

Protocols

Protein Expression Functional Test

Records

Tyrosinase activity assay for 4,8,12 hours incubation with arabinose at 5 different concentrations. We cultured them at 37 degree this time.

Tyrosinase activity assay reagents set-up:

Sample Background Control (SBC) mix: We use only one SBC for each time point incubation of protein samples as we were running out of reagents.

Assay buffer	45ul x 3 = 135ul
Enhancer	5ul x 3 = 15ul

Reaction mix: add this into assay background control, samples and positive control wells

Assay buffer	35ul x 21 = 735ul
Substrate	10ul x 21 = 210ul
Enhancer	5ul x 21 = 105ul

Positive control well: 2ul positive control + 48ul assay buffer

Assay background control: 50ul Assay buffer

Plates:

SDS-page gel loading sequence:

First gel (OPHT-adhesion system):

Ladder- Ladder- Control- OPHT1 - OPHT2 - OPHT3 - OPHM - OPHA1

Second gel (ftNA):

Ladder - Control - ftnA1.1 - ftnA1.2 - ftnA1.3 - ftnA2.1 - ftnA2.2 - ftnA2.3

Results

In the functional test results page

Performers	Philip, Renee
Experiments	ftnA functional test
Protocols	Functional Test
Records	Centrifuged the second bench ftnA test samples and resuspend in LB broth. Measure the OD value. Begin magnet attraction test at 9:30 and will check their states every hour. Transformation of pBAD24-FtnA #2 into seven strains which are Stbl3, DH5 alpha, BL-21 DE3, BL-21 Star, BL-21 Rosetta, BL-21 pLys, TOP10. Started incubation at 13:00. Reset pBAD24-FtnA 1.1, 1.2, 2.1, 2.2 attraction test with only one magnet under it at 14:50. Induced 5 tubes 5ml bacteria pBAD24-FtnA 1.1 with 1ml 100mM ferric ammonium citrate at 16:20. 101219: Incubated 20 mL BL-21 NSP4 tyrosinase on 37 degree for adhesion system functional test with our own enzyme.
Results	None

Performers	Philip
Experiments	Adhesion system functional test
Protocols	Functional Test
Records	Started adhesion system functional test. Incubated adhesion system bacteria with yellow nanoparticle. Would collect the samples at 4 different time points, 1 hr, 2 hr, 3 hr and 4 hr and compare the difference in the relative fluorescence.
Results	None

Performers	Philip, Julien
Experiments	Bacteria culture, Incubate with antibody for immunoblotting membrane
Protocols	Plasmid Construction Protein Expression
Records	Incubated 5ml culture of FtnA-Stbl3,Dh5alpha,BL-21(De3),BL-21(star), BL-21(plys), TOP10, BL-21(Rosetta), And Bl21(WT) at 37 degree shaking incubator started 14:39. Pinto made SDS gel and store in Buffer. Measured protein concentration of FtnA protein samples and stored in -80C. Autoclaved some tips blue and yellow tips and 1.5 mL centrifuge tubes. Incubated FtnA samples and RpoB internal control with anti-flag and anti-rabbit primary antibody. Put 5 mL and 3 mL respectively of the antibodies. Put in 4 degree in Henry’s fridge at 20:20. Incubated 6 20mL culture of OPHT, OPHT2,OPHT3, OPHM, OPHA and WT bl-21 all picked from glycerol stock colony #1 in 37 degree shaking incubator overnight(start 21:00) Has white tape mark, will induce it with 1mM IPTG for 4 hrs after 12 hrs (9:00). For time dependent functional test with yellow fluorescent nanoparticle and Professor Xuan Jun’s nanoparticle. Incubated 5x 20 mL culture of NSP4_BL21 subcultured from yesterday’s overnight culture(1:100 ratio) at 37C started at (21:00) . Will induce later with 0.2% arabinose after 3hrs (00:00). And let it produce tyrosinase into the supernatant for overnight(12-16 hrs) Has green tape mark. This is for testing our own secreted tyrosinase whether it would be able to activate our capture system bacteria. Incubated 6x 5mL tubes of OPHT,OPHT2,OPHT3,OPHM, OPHA and negative ctrl at 37 C has yellow tape mark started (21:00). Picked from glycerol stock colony #1. Will incubate overnight and induce tomorrow after 16 hrs with IPTG and incubate in 30C for 4 hrs (12:00). Incubated 7x 5mL tubes for each bacteria strains we had cultured overnight FtnA-Stbl3,Dh5alpha,BL-21(De3),BL-21(star), BL-21(plys), TOP10, BL-21(Rosetta), And Bl21(WT) at 37 degree shaking incubator started 21:00. Has red tape. Subcultured 1:100 ratio will induce with 0.2% arabinose later after 3 hrs (00:00) and incubate for 12-16 hrs. These samples are for protein concentration check for FtnA based on different strains. Will extract proteins tomorrow and measure protein concentration along with the preparation of protein samples for western blot running on Wednesday. Induced earlier log picked strains incubated this morning from 2:39 5ml cultures of FtnA-Stbl3,Dh5alpha,BL-21(De3),BL-21(star), BL-21(plys), TOP10, BL-21(Rosetta), And Bl21(WT) with 0.2% arabinose (50 ul of 20% arabinose) and stored in 37 degree shaking incubator has purple tape. Started at 21:42. Will incubate overnight for 12 hrs. Then would put with ~20 mM ammonium ferric Citrate 1 mL each and incubate for 24 hrs for magnetization test on Wednesday morning 9:00 am.
Results	None

Performers	Philip, Julien
Experiments	Western blot
Protocols	Protein Expression
Records	Induced red and green taped samples from previous logs with 0.2% arabinose at 00:44 and put in the 37 C shaking incubator. Plan to incubate for 12-16 hrs (Probably 16 hrs). Induced yellow tape marked tubes and white marked tape tubes with 1mM IPTG at 12:00, put them in 30C shaking incubator of Professor Xu's lab. Washed the membrane with tbst 3 times and incubated with anti-rabbit 2nd antibody at 12.40pm
Results	None

Performers	Philip, Julien
Experiments	Western blot
Protocols	Protein Expression
Records	Incubated purple taped samples from 101419 with ~20 mM ferric ammonium citrate. At 37 C starting 00:49. Run SDS-Page gel. Loading order as below: First gel Ladder-Ladder-Control-2%-0.2%-0.02%-0.002%-0.0002% Second gel Ladder-BL-21WT-Rosetta-BL-21(star)-TOP10-BL-21(plys)-Dh5alpha-BL-21(De3) Incubated membranes with primary antibody for overnight. (Anti-flag for 19kda ftna, Anti-rpob for 150kda internal ctrl).
Results	None

Performers	Philip
Experiments	Immunofluorescence
Protocols	Protein Expression
Records	Incubated immunofluorescence slide samples with histidine primary antibody for 12-16 hrs. Put in 4 degree in Henry’s fridge.
Results	None

Performers	Philip, Renee
Experiments	Cell viability testing
Protocols	Functional Test
Records	Started incubation of bacteria cell viability with ZnO and AgNP at 04:30. Will test the following time points (Hrs) or OD600 (2, 4,6,8,10,12,14,16). 6:30, 8:30, 10:30, 12:30, 14:30, 16:30, 18:30 are the time points. I would need help getting the absorbance values for the 10:30, 12:30, 14:30 timepoints. Incubated 20 mL overnight culture OPHt,OpHT2, OPHT3, OPHM, OPHA, -ctrl in 37 C shaking incubator started 17:00.
Results	None

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-       <div class="April"><button class="modal-open" data-modal="modal3">27</button></div>
+       <div class="May"><button class="modal-open" data-modal="modal0525">25</button></div>
-       <div class="April"><button class="modal-open" data-modal="modal1">28</button></div>
+       <div class="May"><button class="modal-open" data-modal="modal0526">26</button></div>
-       <div class="April"><button class="modal-open" data-modal="modal2">29</button></div>
+       <div class="May"><button class="modal-open" data-modal="modal0527">27</button></div>
-       <div class="April"><button class="modal-open" data-modal="modal3">30</button></div>
+      <div><button disabled class="modal-open" data-modal="modal0528">28</button></div>
+       <div class="May"><button class="modal-open" data-modal="modal0529">29</button></div>
+       <div class="May"><button class="modal-open" data-modal="modal0530">30</button></div>
+       <div class="May"><button class="modal-open" data-modal="modal0531">31</button></div>
      </div>
    </div>
    </div>
 </div>
-<!--End of April Calendar-->
+<!--End of May Calendar-->
-<!--April Modals-->
+<!--May Modal-->
-   <div class="modal" id="modal2">
+   <div class="modal" id="modal0520">
      <div class="modal-content">
-       <div class="modal-header">Modal 2
+       <div class="modal-header">May 20, 2019
          <span class="modal-close closeicon">&times;</span></div>
-       <div class="modal-body">Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. </div>
+       <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>gBlocks PCR amplification: IDT-OMPHT,NSP4-tyrosinase,Mefp-5</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>PCR reaction: Prepare reaction mix on ice. Primers for IDT-OMPHT are 100-F/R, for NSP4 tyrosinase are 113/114-F/R, for Mefp-5 are 105-F/R. Concentrations: IDT-OMPHT is 11.0 ng/μl, NSP4-Tyrosinase is 16.7 ng/μl, Mefp-5 is 18.2 ng/μl.</p>
+            <br>
+<!--Table Template-->
+            <table>
+            <tr>
+            <td>Q5® High-Fidelity 2X Master Mix</td>
+            <td>25 ul</td>
+            </tr>
+            <tr>
+              <td>Primer mix (10cM)</td>
+              <td>2.5ul</td>
+            </tr>
+            <tr>
+              <td>Template (IDT-OMPHT/NSP4 tyrosinase/Mefp-5)</td>
+              <td>Xμl
+              <p>(We used 10 ng of the fragment as template for each of them)</p></td>
+            </tr>
+            <tr>
+              <td>ddH20</td>
+              <td>22.5-X</td>
+            </tr>
+            <tr>
+              <td>Total</td>
+              <td>50μl</td>
+            </tr>
+            </table>
+<!--End of Table-->
+            <p>PCR setting: Tm for IDT-OMPHT is 68°C, for NSP4 tyrosinase is 62°C, for Mefp-5 is 65°C.</p>
+<table width="495">
+<tbody>
+<tr>
+<td width="168">
+<p>Initial denaturation</p>
+</td>
+<td width="158">
+<p>98&deg;C</p>
+</td>
+<td width="169">
+<p>30s</p>
+</td>
+</tr>
+<tr>
+<td rowspan="3" width="168">
+<p>X35 Cycles</p>
+</td>
+<td width="158">
+<p>98&deg;C</p>
+</td>
+<td width="169">
+<p>10s</p>
+</td>
+</tr>
+<tr>
+<td width="158">
+<p>Tm</p>
+</td>
+<td width="169">
+<p>20s</p>
+</td>
+</tr>
+<tr>
+<td width="158">
+<p>72&deg;C</p>
+</td>
+<td width="169">
+<p>45s</p>
+</td>
+</tr>
+<tr>
+<td width="168">
+<p>Final extension</p>
+</td>
+<td width="158">
+<p>72&deg;C</p>
+</td>
+<td width="169">
+<p>2min</p>
+</td>
+</tr>
+<tr>
+<td width="168">
+<p>Hold</p>
+</td>
+<td width="158">
+<p>4&deg;C</p>
+</td>
+<td width="169">
+<p>Infinite</p>
+</td>
+</tr>
+</tbody>
+</table>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
        <div class="modal-footer"><button class="link modal-close">Close</button></div>
      </div>
    </div>
+<!--End of Modals-->
-   <div class="modal" id="modal1">
+<!--Modal Template-->
+   <div class="modal" id="modal0521">
      <div class="modal-content">
-       <div class="modal-header">Modal 1
+       <div class="modal-header">May 21, 2019
          <span class="modal-close closeicon">&times;</span></div>
-       <div class="modal-body">Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. </div>
+       <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>052019 gBlocks PCR fragments checking via gel running</p>
+                <p>Repeat gBlocks PCR amplification: IDT-OMPHT,NSP4 tyrosinase,Mefp-5</p>
+                <p>052191 gBlocks PCR fragments checking via gel running</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Gel running: Make two different size gels. The small one is 25ml TAE with 0.25g agarose and 1.25ml SYBR safe dye. This gel is for checking the 052019 PCR fragments. The big one is 50ml TAE with 0.5g agarose and 2.5ml SYBR safe dye. This gel is for checking the 052119 PCR fragments. Mix 5μl PCR product and 1μl 6X loading dye. Run at 130V for 30 min and then check the image.</p>
+            <br>
+            <p>PCR reaction and setting: The reaction mix is the same as 052019. Only difference we made is that we changed the Tm into three sets in order to figure out whether Tm led to the incorrect bands for image052019. Tm for Set A is 65°C, Tm for set B is 57°C, and Tm for Set C is 53°C.</p>
+            <br>
+            <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/2/26/T--UM_Macau--PicUploadTesting.png">
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>After rechecking, we found that we mistakenly ordered the primers for NSP4 tyrosinase. Actually the fragment we ordered has already contained the homologous regions that we wanted.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
        <div class="modal-footer"><button class="link modal-close">Close</button></div>
      </div>
    </div>
+<!--End of Modals-->
-   <div class="modal" id="modal3">
+<!--Modal Template-->
+   <div class="modal" id="modal0523">
      <div class="modal-content">
-       <div class="modal-header">Modal 3
+       <div class="modal-header">May 23, 2019
          <span class="modal-close closeicon">&times;</span></div>
-       <div class="modal-body">Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat.
+       <div class="modal-body">
-         <img src="home/T--UM_Macau--home1.jpg" style="width: 100%"></div>
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Restriction digestion of plasmid vectors</p>
+                <p>Recheck gBlocks NSP4 tyrosinase</p></td>
+          </tr>
+        <!--Protocols-->
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Digestion reaction:</p>
+<table style="height: 238px;" width="322">
+<tbody>
+<tr>
+<td style="width: 156px;">
+<p>pBAD24 (102.9 ng/&mu;l)</p>
+</td>
+<td style="width: 150px;">
+<p>30 &mu;l (30mg of DNA)</p>
+</td>
+</tr>
+<tr>
+<td style="width: 156px;">
+<p>EcoRI-HF</p>
+</td>
+<td style="width: 150px;">
+<p>1 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td style="width: 156px;">
+<p>XbaI</p>
+</td>
+<td style="width: 150px;">
+<p>1 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td style="width: 156px;">
+<p>Cut smart</p>
+</td>
+<td style="width: 150px;">
+<p>5 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td style="width: 156px;">
+<p>ddH2O</p>
+</td>
+<td style="width: 150px;">
+<p>13 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td style="width: 156px;">
+<p>Total</p>
+</td>
+<td style="width: 150px;">
+<p>50&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+            <p>Incubate at 37°C overnight</p>
+            <br>
+            <p>Gel running: Run the PCR products of IDT-OMPHT and Mefp-5, and directly run the gBlocks NSP4 tyrosinase at 130V for 30 min and then check the image.</p>
+          </td>
+            </td>
+          </tr>
+         <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/b/b0/T--UM_Macau--Lablog_23_May.png" style="width:50%">
+              <p>We got the correct band for NSP4 tyrosinase even though it was very weak. We would try to amplify after ligation.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
        <div class="modal-footer"><button class="link modal-close">Close</button></div>
      </div>
    </div>
+<!--End of Modals-->
-<!--End of April Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0524">
+    <div class="modal-content">
+      <div class="modal-header">May 24, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Gel running:&nbsp;50 &mu;l digested sample + 10 &mu;l loading dye. The order of loading is shown below. Then separate 60&mu;l into two wells on the gel because the well will be overflowing if we put all 60 &mu;l into one well.</p>
+<table width="530">
+<tbody>
+<tr>
+<td width="113">
+<p>1 kb ladder</p>
+<p>(2 &mu;l)</p>
+</td>
+<td width="142">
+<p>Sample</p>
+<p>(30 &mu;l)</p>
+</td>
+<td width="151">
+<p>Sample</p>
+<p>(less than 30 &mu;l)</p>
+</td>
+<td width="124">
+<p>1 kb ladder</p>
+<p>(2 &mu;l)</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Run at 120V for 23 min in order to separate the digestion products and obtain the target vector (pBAD24, 4516 bp) from unused fragments (26 bp).</p>
+<p>Gel purification: We&rsquo;re using protocol from Illustra GFX PCR DNA and Gel Band Purification Kit Gel. Gel slices weight are 258 mg (used 258 &mu;l CBT3) and 280 mg (used 280 &mu;l CBT3)*CBT3 = capture buffer type 3</p>
+<p>Sample capture &rarr; sample binding &rarr; wash &amp; dry &rarr; elution&nbsp;</p>
+<p>We used 1 Microspin column and 1 collection tube for purification of 2 gels. Then Elution with 20&mu;l elution buffer type 6&nbsp;</p>
+<p>Gibson Assembly reagents calculation:</p>
+<table width="625">
+<tbody>
+<tr>
+<td width="313">
+<p>Insert (NSP4 + tyrosinase)</p>
+</td>
+<td width="313">
+<p>Vector (pBAD24)</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Length: 1004 bp</p>
+</td>
+<td width="313">
+<p>Length: 4542 bp</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Concentration: 16.7 ng/&mu;l</p>
+</td>
+<td width="313">
+<p>Concentration: 32.80 ng/&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Volume used for ligation: 5 &mu;l</p>
+</td>
+<td width="313">
+<p>Volume used for ligation: 3 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Mass: 83.5 ng</p>
+</td>
+<td width="313">
+<p>Mass: 98.4 ng</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Mole: 134.6 fmol / 0.1346 pmol</p>
+<p>(by NEBioCalculator)</p>
+</td>
+<td width="313">
+<p>Mole: 35.06 fmol / 0.03506 pmol</p>
+<p>(by NEBioCalculator)</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Volume used (Vector : Insert = 1 : 4)</p>
+<p>Gibson assembly reaction:</p>
+<table width="378">
+<tbody>
+<tr>
+<td width="313">
+<p>Insert (NSP4 + tyrosinase)</p>
+</td>
+<td width="65">
+<p>5 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Vector (pBAD24)</p>
+</td>
+<td width="65">
+<p>3 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="65">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Deionized H2O</p>
+</td>
+<td width="65">
+<p>2 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Total</p>
+</td>
+<td width="65">
+<p>20 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
-<!--May Calendar-->
-<div id="May" class="vertabcontent">
-  <div class="wrapper">
-  <div class="calendar">
-    <div class="month" style="background-color: #d53b48">
-      <div>
-        <h1>May</h1>
        </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
      </div>
+  </div>
+<!--End of Modals-->
-    <div class="weekends" style="background-color: #b22a35">
+<!--Modal Template-->
-       <div>Sun</div>
+  <div class="modal" id="modal0525">
-       <div>Mon</div>
+    <div class="modal-content">
-      <div>Tue</div>
+       <div class="modal-header">May 25, 2019
-      <div>Wed</div>
+        <span class="modal-close closeicon">&times;</span></div>
-      <div>Thu</div>
+       <div class="modal-body">
-      <div>Fri</div>
-       <div>Sat</div>
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Gibson assembly reaction as negative control for 052419:</p>
+<table width="226">
+<tbody>
+<tr>
+<td width="151">
+<p>Master Mix</p>
+</td>
+<td width="76">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="151">
+<p>Digested Plasmid</p>
+</td>
+<td width="76">
+<p>2 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="151">
+<p>ddH2O</p>
+</td>
+<td width="76">
+<p>18 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="151">
+<p>Total</p>
+</td>
+<td width="76">
+<p>30 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Incubated at 50&deg;C for 15 minutes.</p>
+<p>&nbsp;</p>
+<p>Transformation of NSP4-Tyrosinase-pBAD24 into competent cells:</p>
+<p>a. 50 &mu;l competent cells + 5 &mu;l Gibson assembly mix made on 052419</p>
+<p>b. 50 &mu;l competent cells + 5 &mu;l negative control made on 052519</p>
+<p>c. Spread on two plates with Ampicillin and incubate at 37&deg;C overnight</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+       <div class="modal-footer"><button class="link modal-close">Close</button></div>
      </div>
+  </div>
+<!--End of Modals-->
-    <div class="days">
+<!--Modal Template-->
-      <div><button disabled class="prev_date">28</button></div>
+  <div class="modal" id="modal0526">
-      <div><button disabled class="prev_date">29</button></div>
+    <div class="modal-content">
-      <div><button disabled class="prev_date">30</button></div>
+       <div class="modal-header">May 26, 2019
-       <div class="May"><button class="modal-open" data-modal="modal1">1</button></div>
+        <span class="modal-close closeicon">&times;</span></div>
-       <div class="May"><button class="modal-open" data-modal="modal2">2</button></div>
+       <div class="modal-body">
-      <div class="May"><button class="modal-open" data-modal="modal3">3</button></div>
-      <div class="May"><button class="modal-open" data-modal="modal1">4</button></div>
+        <table>
-      <div class="May"><button class="modal-open" data-modal="modal2">5</button></div>
-      <div class="May"><button class="modal-open" data-modal="modal3">6</button></div>
+        <!--Experiments-->
-      <div class="May"><button class="modal-open" data-modal="modal1">7</button></div>
+          <tr>
-      <div class="May"><button class="modal-open" data-modal="modal2">8</button></div>
+            <th>Experiments</th>
-      <div class="May"><button class="modal-open" data-modal="modal3">9</button></div>
+            <td><p>Tyrosinase secretion system construction</p></td>
-      <div class="May"><button class="modal-open" data-modal="modal1">10</button></div>
+          </tr>
-      <div class="May"><button class="modal-open" data-modal="modal2">11</button></div>
-      <div class="May"><button class="modal-open" data-modal="modal3">12</button></div>
+        <!--Protocols-->
-      <div class="May"><button class="modal-open" data-modal="modal1">13</button></div>
+          <tr>
-      <div class="May"><button class="modal-open" data-modal="modal2">14</button></div>
+            <th>Protocols</th>
-      <div class="May"><button class="modal-open" data-modal="modal3">15</button></div>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
-      <div class="May"><button class="modal-open" data-modal="modal1">16</button></div>
+          </tr>
-      <div class="May"><button class="modal-open" data-modal="modal2">17</button></div>
-      <div class="May"><button class="modal-open" data-modal="modal3">18</button></div>
+        <!--Records-->
-      <div class="May"><button class="modal-open" data-modal="modal1">19</button></div>
+          <tr>
-      <div class="May"><button class="modal-open" data-modal="modal2">20</button></div>
+            <th>Records</th>
-      <div class="May"><button class="modal-open" data-modal="modal3">21</button></div>
+            <td><p>Colony picking and incubation:</p>
-      <div class="May"><button class="modal-open" data-modal="modal1">22</button></div>
+<table style="width: 378px;">
-      <div class="May"><button class="modal-open" data-modal="modal2">23</button></div>
+<tbody>
-      <div class="May"><button class="modal-open" data-modal="modal3">24</button></div>
+<tr>
-      <div class="May"><button class="modal-open" data-modal="modal1">25</button></div>
+<td style="width: 192px;">
-      <div class="May"><button class="modal-open" data-modal="modal2">26</button></div>
+<p>Plate</p>
-      <div class="May"><button class="modal-open" data-modal="modal3">27</button></div>
+</td>
-      <div class="May"><button class="modal-open" data-modal="modal1">28</button></div>
+<td style="width: 172px;">
-      <div class="May"><button class="modal-open" data-modal="modal2">29</button></div>
+<p>Colony number</p>
-       <div class="May"><button class="modal-open" data-modal="modal3">30</button></div>
+</td>
-       <div class="May"><button class="modal-open" data-modal="modal3">31</button></div>
+</tr>
+<tr>
+<td style="width: 192px;">
+<p>Negative control</p>
+</td>
+<td style="width: 172px;">
+<p>9 small colonies</p>
+</td>
+</tr>
+<tr>
+<td style="width: 192px;">
+<p>pBAD24-NSP4 tyrosinase</p>
+</td>
+<td style="width: 172px;">
+<p>1 colony</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Prepare the LB Broth with Ampicillin antibiotic and tubes with 1 mL &ndash; 5 mL LB Broth with Ampicillin antibiotic.&nbsp;</p>
+<p>Pick the only one single colony from the pBAF24-NSP4 tyrosinase transformation plate.</p>
+<p>And randomly pick 5 colonies from the negative control transformation plate. Incubate at 37&deg;C overnight.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/3/3d/T--UM_Macau--Lablog_26_May_1.png" style="width: 40%;">
+            <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/5/57/T--UM_Macau--Lablog_26_May_2.png" style="width: 40%;">
+            <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/1/1d/T--UM_Macau--Lablog_26_May_3.png" style="width: 80%; margin-top:5px">
+              <p>The negative control has much more colonies than our pBAD24-NSP4 tyrosinase. So we guessed we might mistakenly record the two plates. The results would show us whether our assumption was true after the plasmid extraction and gel running.</p>
+            </td>
+          </tr>
+        </table>
+       </div>
+       <div class="modal-footer"><button class="link modal-close">Close</button></div>
      </div>
    </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0527">
+    <div class="modal-content">
+      <div class="modal-header">May 27, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Plasmid extraction:</p>
+<p>a. Collect 3 mL of overnight suspension culture from the five negative control tubes.</p>
+<p>b. Elute with 50&mu;l elution buffer. Did the elution steps three times with the same 50&mu;l elution buffer. Incubate 4min between each centrifuge.</p>
+<p>c. Then use Nanodrop to test the extracted plasmid concentration.&nbsp;</p>
+<p>&nbsp;</p>
+<p>Restriction enzyme digestion reaction: Total 50&mu;l</p>
+<table width="516">
+<tbody>
+<tr>
+<td width="123">
+<p>DNA (500 ng)</p>
+</td>
+<td width="72">
+<p>39.1&mu;l</p>
+</td>
+<td width="76">
+<p>27.2&mu;l</p>
+</td>
+<td width="85">
+<p>16&mu;l</p>
+</td>
+<td width="76">
+<p>18&mu;l</p>
+</td>
+<td width="85">
+<p>18&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>EcoRI-HF</p>
+</td>
+<td width="72">
+<p>0.5&mu;l</p>
+</td>
+<td width="76">
+<p>0.5&mu;l</p>
+</td>
+<td width="85">
+<p>0.5&mu;l</p>
+</td>
+<td width="76">
+<p>0.5&mu;l</p>
+</td>
+<td width="85">
+<p>0.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>XbaI</p>
+</td>
+<td width="72">
+<p>0.5&mu;l</p>
+</td>
+<td width="76">
+<p>0.5&mu;l</p>
+</td>
+<td width="85">
+<p>0.5&mu;l</p>
+</td>
+<td width="76">
+<p>0.5&mu;l</p>
+</td>
+<td width="85">
+<p>0.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>Cut smart</p>
+</td>
+<td width="72">
+<p>5&mu;l</p>
+</td>
+<td width="76">
+<p>5&mu;l</p>
+</td>
+<td width="85">
+<p>5&mu;l</p>
+</td>
+<td width="76">
+<p>5&mu;l</p>
+</td>
+<td width="85">
+<p>5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>ddH2O</p>
+</td>
+<td width="72">
+<p>4.9&mu;l</p>
+</td>
+<td width="76">
+<p>16. 8&mu;l</p>
+</td>
+<td width="85">
+<p>28&mu;l</p>
+</td>
+<td width="76">
+<p>26&mu;l</p>
+</td>
+<td width="85">
+<p>26&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Incubate at 37&deg;C overnight</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>DNA plasmid extraction: They are 12.81ng/μl, 18.38ng/μl, 31.41ng/μl, 27.2ng/μl and 28.44ng/μl. </p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
    </div>
-</div>
+<!--End of Modals-->
-<!--End of May Calendar-->
+<!--Modal Template-->
+  <div class="modal" id="modal0529">
+    <div class="modal-content">
+      <div class="modal-header">May 29, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Plasmid identity checking for 052619: Run the digested plasmid from five randomly picked colonies on 052619. Loading order for the gel: Left to right 1 kb ladder -> Colony 1 -> Colony 2 -> Colony 3 -> Colony 4 -> Colony 5</p>
+            <br>
+            <p>Repeat transformation of bacteria: 5 μl Gibson assembly mix made on 052419 and 5 μl negative control made on 052519. Spread on two plates with Ampicillin and incubate at 37°C overnight.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+            <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/d/d4/T--UM_Macau--Lablog_29_May.jpg" style="width: 60%; display:block; margin-left: auto; margin-right: auto;">
+              <p>Besides the result of colony 3 that we don’t know how to explain it. Its band is neither the vector nor the NSP4 tyrosinase which was 1022 bp, but this band is around 1500 bp. All the other bands are our vectors which is pBAD24. However, they do not have the insert so they are not the proper plasmid.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0530">
+    <div class="modal-content">
+      <div class="modal-header">May 30, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Colony picking and incubation</p>
+<table width="380">
+<tbody>
+<tr>
+<td width="218">
+<p>Plate</p>
+</td>
+<td width="163">
+<p>Colony number</p>
+</td>
+</tr>
+<tr>
+<td width="218">
+<p>Negative control</p>
+</td>
+<td width="163">
+<p>3 small colonies</p>
+</td>
+</tr>
+<tr>
+<td width="218">
+<p>pBAD24-NSP4 tyrosinase</p>
+</td>
+<td width="163">
+<p>2 colonies</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Pick the two colonies from the pBAF24-NSP4 tyrosinase transformation plate.</p>
+<p>And pick the 3 colonies from the negative control transformation plate. Incubate at 37&deg;C overnight.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+            <div class="row" style="width:80%; margin-top:5px">
+              <div class="col-sm-6">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/a/a8/T--UM_Macau--Lablog_30_May_1.png" >
+              </div>
+              <div class="col-sm-6">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/d/de/T--UM_Macau--Lablog_30_May_2.png">
+              </div>
+            </div>
+            <div class="row" style="width:80%; margin-top:5px">
+              <div class="col-sm-6">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/8/8c/T--UM_Macau--Lablog_30_May_3.png" >
+              </div>
+              <div class="col-sm-6">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/b/bb/T--UM_Macau--Lablog_30_May_4.png">
+              </div>
+            </div>
+            <div class="row" style="width:80%; margin-top:5px">
+              <div class="col-sm-6">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/0/05/T--UM_Macau--Lablog_30_May_5.png" >
+              </div>
+              <div class="col-sm-6">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/3/3e/T--UM_Macau--Lablog_30_May_6.png">
+              </div>
+            </div>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0531">
+    <div class="modal-content">
+      <div class="modal-header">May 31, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Plasmid extraction:</p>
+<p>a. Collect 3 ml of overnight suspension culture from the two pBAD24-NSP4 tyrosinase tubes.</p>
+<p>b. Elute with 50&mu;l elution buffer. Did the elution steps three times with the same 50&mu;l elution buffer. Incubate 4min between each centrifuge.</p>
+<p>c. Then use Nanodrop to test the extracted plasmid concentration. They are 46ng/&mu;l and 28ng/&mu;l.</p>
+<p>&nbsp;</p>
+<p>Restriction enzyme digestion reaction: Total 25&mu;l</p>
+<table width="270">
+<tbody>
+<tr>
+<td width="123">
+<p>DNA (500 ng)</p>
+</td>
+<td width="72">
+<p>15&mu;l</p>
+</td>
+<td width="76">
+<p>18&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>EcoRI-HF</p>
+</td>
+<td width="72">
+<p>0.5&mu;l</p>
+</td>
+<td width="76">
+<p>0.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>XbaI</p>
+</td>
+<td width="72">
+<p>0.5&mu;l</p>
+</td>
+<td width="76">
+<p>0.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>Cut smart</p>
+</td>
+<td width="72">
+<p>2.5&mu;l</p>
+</td>
+<td width="76">
+<p>2.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="123">
+<p>ddH2O</p>
+</td>
+<td width="72">
+<p>10.5&mu;l</p>
+</td>
+<td width="76">
+<p>3.5&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Incubate at 37&deg;C overnight</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of May Modals-->
+<!--June Calendar-->
 <div id="June" class="vertabcontent">
    <div class="wrapper">
    <div class="calendar">
-     <div class="month" style="background-color:#26d8d2">
+     <div class="month" style="background-color:#7460a4">
        <div>
          <h1>June</h1>
@@ Line 796: / Line 1,802: @@
      </div>
-     <div class="weekends" style="background-color:#20b2ad">
+     <div class="weekends" style="background-color:#584486">
        <div>Sun</div>
        <div>Mon</div>
@@ Line 813: / Line 1,819: @@
        <div><button disabled class="prev_date">30</button></div>
        <div><button disabled class="prev_date">31</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal1">1</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0601">1</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal2">2</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0602">2</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal3">3</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0603">3</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal1">4</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0604">4</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal2">5</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0605">5</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal3">6</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0606">6</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal1">7</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0607">7</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal2">8</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0608">8</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal3">9</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0609">9</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0611">11</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0612">12</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0613">13</button></div>
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+       <div class="June"><button class="modal-open" data-modal="modal0614">14</button></div>
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+       <div class="June"><button class="modal-open" data-modal="modal0615">15</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal1">16</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0616">16</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0617">17</button></div>
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+       <div class="June"><button class="modal-open" data-modal="modal0618">18</button></div>
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+       <div class="June"><button class="modal-open" data-modal="modal0619">19</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal2">20</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0620">20</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal3">21</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0621">21</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0622">22</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0623">23</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0624">24</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0625">25</button></div>
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+       <div class="June"><button class="modal-open" data-modal="modal0626">26</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal3">27</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0627">27</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal1">28</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0628">28</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal2">29</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0629">29</button></div>
-       <div class="June"><button class="modal-open" data-modal="modal3">30</button></div>
+       <div class="June"><button class="modal-open" data-modal="modal0630">30</button></div>
      </div>
    </div>
    </div>
 </div>
+<!--End of June Calendar-->
+<!--Modal 0601-->
+  <div class="modal" id="modal0601">
+    <div class="modal-content">
+      <div class="modal-header">June 1, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase secretion system construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Plasmid identity checking for 053019: Run the digested plasmid from two picked colonies from pBAD24-NSP4 tyrosinase plate on 053019 at 120V for 25 min. Loading order is 1 kb ladder, sample1, sample2, 1 kb ladder. Load 25μl digestion product with 5μl loading dye.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/8/8d/T--UM_Macau--Lablog_1_Jun.jpg" style="width:60%">
+              <p>No correct colonies. Through these two transformation experiments, we thought our competent cells have some problem. So we would change to use a new one.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0601-->
+<!--Modal0603-->
+  <div class="modal" id="modal0603">
+    <div class="modal-content">
+      <div class="modal-header">June 3, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Vector identities checking via gel running</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Expected band size for each plasmid backbone:</p>
+<p>pBBR1MCS2 = 5148 bp and pBAD24 = 4542 bp</p>
+<br>
+<p>I: First gel running around 10 am by Hugh</p>
+<p>Samples loaded&nbsp;:</p>
+<p>10&mu;l pBBR1MCS-2 + 2&mu;l loading dye (x6)</p>
+<p>10&mu;l pBAD24 + 2&mu;l loading dye (x6)</p>
+<br>
+<p>Ran for 120 V for 25 minutes. Initial observation was that the samples were floating and not settling down even after the addition of the loading dye. Hugh did not run this gel and prepared a new one instead because the TA said maybe the gel preparation is not good.</p>
+<br>
+<p>II: Second gel running around 11 am by Hugh</p>
+<p>Samples loaded: </p>
+<p>10&mu;l pBBR1MCS-2 + 2&mu;l loading dye (x6)</p>
+<p>10&mu;l pBAD24 + 2&mu;l loading dye (x6)</p>
+<br>
+<p>Ran for 120 V for 25 minutes. Gel image is figure 1.</p>
+<br>
+<p>III: Third gel running around 5 pm by Philip</p>
+<p>Samples loaded: </p>
+<p>24&mu;l pBBR1MCS-2 + 20&mu;l loading dye (x6)</p>
+<p>24&mu;l pBAD24 + 20&mu;l loading dye (x6)</p>
+<br>
+<p>Ran for 120 V for 25 minutes. Initial observation: Even after mixing the large volume of loading dye. When loading the sample it was still floating away. We now suspected that there might be something wrong with the sample. We think the sample contains ethanol.&nbsp;Gel image is figure 2.</p>
+<br>
+<p>IV: Fourth gel running around 6 pm by Philip</p>
+<p>Samples loaded:</p>
+<p>10&mu;l pBBR1MCS-2 + 2&mu;l loading dye (x6)</p>
+<p>10&mu;l pBAD24 + 2&mu;l loading dye (x6)</p>
+<br>
+<p>Ran for 120 V for 25 minutes. Initial observation: Same thing as the previous gel</p>
+<br>
+<p>No correct band at all, I think it&rsquo;s due to the loading dye making the fragments too heavy and thus it did not run fast in the gel, therefore our Xiao Xiong (One of our TAs)&nbsp; recommended to prepare a new gel and use a smaller well size. After this result, Xiao Xiong has put the stock plasmid backbone tubes onto a heating block; she opened the cap of the tubes and heated the tubes at 55 C, I reclaimed the stuff after maybe 30 minutes; this was to evaporate possible ethanol contaminants.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/1/11/T--UM_Macau--Lablog_3_Jun_1.png" style="width: 50%">
+              <p>Sample might have floated away and none were actually run in the gel.</p>
+              <br>
+              <br>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/d/d3/T--UM_Macau--Lablog_3_Jun_2.png" style="width: 50%">
+              <p>No correct band at all, I think it’s due to the loading dye making the fragments too heavy and thus it did not run fast in the gel, therefore our Xiao Xiong (One of our TAs)  recommended to prepare a new gel and use a smaller well size. After this result, Xiao Xiong has put the stock plasmid backbone tubes onto a heating block; she opened the cap of the tubes and heated the tubes at 55 C, I reclaimed the stuff after maybe 30 minutes; this was to evaporate possible ethanol contaminants.</p>
+              <br>
+              <br>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/8/83/T--UM_Macau--Lablog_3_Jun_3.png" style="width: 50%">
+              <p>It seems that the band is still not at the correct place. There is not even a trace of the correct band for pBAD24 it seems like the band is located more than 10kb. For pBBR1MCS-2, it seems like the DNA sample all floated away and what we saw that settled were probably just the loading dye. Next time we should mix the loading dye and the DNA sample better. </p>
+              <br>
+              <br>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/9/97/T--UM_Macau--Lablog_3_Jun_4.jpg" style="width: 50%">
+              <p>Maybe there is something wrong with the plasmid? After consultation with Professor Zheng Jun’s PhD. Student, apparently we should first digest the plasmid vector with at least one enzyme to linearize it. Only then should we run it in the gel and check the size. Apparently if we do not linearize the plasmid before running, due to its circular form it would run slower along the gel causing it to show a non-true band.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0603-->
+<!--Modal0604-->
+  <div class="modal" id="modal0604">
+    <div class="modal-content">
+      <div class="modal-header">June 4, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat vector identities checking via gel running</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+              <p>
+                Restriction enzyme digestion reaction for vectors: Total 10μl
+A1 is pBBR1MCS-2 041719, A2 is pBBR1MCS-2 050519, B1 is pBAD24 041719, B2 is pBAD24 050519.
+              </p>
+              <br>
+              <table>
+                <tr>
+                  <td>Vector</td>
+                  <td>A1</td>
+                  <td>A2</td>
+                  <td>A3</td>
+                  <td>A4</td>
+                </tr>
+                <tr>
+                  <td>DNA (500 ng)</td>
+                  <td>4μl</td>
+                  <td>8.5μl</td>
+                  <td>1μl</td>
+                  <td>5μl</td>
+                </tr>
+                <tr>
+                  <td>EcoRI-HF</td>
+                  <td>0.5μl</td>
+                  <td>0.5μl</td>
+                  <td>0.5μl</td>
+                  <td>0.5μl</td>
+                </tr>
+                <tr>
+                  <td>Cut Smart</td>
+                  <td>1μl</td>
+                  <td>1μl</td>
+                  <td>1μl</td>
+                  <td>1μl</td>
+                </tr>
+                <tr>
+                  <td>ddH20</td>
+                  <td>4.5μl</td>
+                  <td>0μl</td>
+                  <td>7.5μl</td>
+                  <td>3.5μl</td>
+                </tr>
+              </table>
+              <br>
+              <p>Incubate at 37℃ for 1h. Gel running: 120V for 25 min. 10μl digestion product + 2μl loading dye. Gel arrangement is 1kb ladder, A1, B1, B2, A2, 1 kb ladder. Restriction enzyme digestion reaction for insert: Total 50μl. Incubate overnight.</p>
+              <br>
+              <table>
+                <tr>
+                  <td>Insert DNA (200 ng)</td>
+                  <td>8.17μl</td>
+                </tr>
+                <tr>
+                  <td>EcoRI-HF</td>
+                  <td>2μl</td>
+                </tr>
+                <tr>
+                  <td>Xbal</td>
+                  <td>2ul</td>
+                </tr>
+                <tr>
+                  <td>ddH20</td>
+                  <td>37.83μl</td>
+                </tr>
+                <tr>
+                  <td>Cut Smart</td>
+                  <td>5μl</td>
+                </tr>
+              </table>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/7/7b/T--UM_Macau--Lablog_4_Jun.jpg" style="width: 60%; margin-bottom: 2%">
+              <br>
+              <p>After digestion with EcoRI-HF for 1 hr at 37 C. We ran the plasmid stock that has kept (041719) along with the stock plasmid solution that has prepared (050519). The 041719 are at the expected size: pBBR1MCS2 = 5148 bp and pBAD24 = 4542 bp.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0604-->
+<!--Modal0605-->
+  <div class="modal" id="modal0605">
+    <div class="modal-content">
+      <div class="modal-header">June 5, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Vector and insert checking via gel running</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+              <p>Sample:55μl digested insert + 10μl loading dye</p>
+              <p>Order: 1 kb ladder(μl), sample(30μl), sample(30μl), 1 kb ladder(2μl)</p>
+              <p>Run at 120V for 30 min. </p>
+              <p>Our expected size is 4515bp for vector pBAD24 and 1004bp for NSP4+tyrosinase</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/7/72/T--UM_Macau--Lablog_5_Jun.jpg" style="width: 80%; margin-bottom:2%">
+              <br>
+              <p>The left one is for vector pBAD24 and the other one is for insert NSP4+tyrosinase. The right one was the supposedly digested insert but we did not see any band there.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0605-->
+<!--Modal0606-->
+  <div class="modal" id="modal0606">
+    <div class="modal-content">
+      <div class="modal-header">June 6, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBAD24, pBBR1MCS2 plasmid extraction for sequencing</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Pellet 4 ml of the 5 ml overnight culture. Pipette 1 ml into microcentrifuge tube and centrifuge for 3 min.&nbsp;Discard supernatant and add next 1 ml into same tube.&nbsp;Repeat until you&rsquo;ve pelleted 4 ml worth of bacteria culture. (12 min of centrifugation)</p>
+<p>Follow protocol in QIAGEN (miniprep) plasmid extraction kit.</p>
+<p>&nbsp;*Miniprep is for 5 ml bacteria culture, yield is low because starting vol. is low.&nbsp;</p>
+<p>&nbsp;&nbsp;&nbsp; Mediprep is for 50 ml, maxi for 100, and giga for 1L.&nbsp;</p>
+<p>P1 is resuspension buffer.&nbsp;</p>
+<p>P2 is lysis buffer. (Alkaline, with many salts)</p>
+<p>N3 is neutralization buffer. (Balance alkalinity)</p>
+<p>You&rsquo;d see some white precipitate after adding N3. The centrifuge step in protocol is to spin down most of that ppt. for you to collect supernatant. This prevents clogging (blocking) of the spin column for downstream procedure.&nbsp;</p>
+<p>Nanodrop at Prof. Xu&rsquo;s Lab. Clean with water and blank, then measure.&nbsp;</p>
+<p>pBAD24: 66.4ng/&mu;l (acceptable)</p>
+<p>pBBR1MCS2: 37.6ng/&mu;l (still pretty low)</p>
+<p>Afternoon: Philip sent them for plasmid sequencing.&nbsp;</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0606-->
+<!--Modal0614-->
+  <div class="modal" id="modal0614">
+    <div class="modal-content">
+      <div class="modal-header">June 14, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Resuspension of gBlock fragments</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Centrifuged the tube at 4500 x g to make sure the dried pellet is at the bottom of the tube. Added 100μl TE buffer to all of the fragments (I was expecting to get 10ng/μl). All products that were delivered were 1000 ng. Vortexed briefly. Incubated at 50 C for 20 minutes. Vortexed then centrifuged. Got the concentration using Nanodrop, we used TE buffer as blank. After the last step, I aliquoted 50μl to another tube so that we would avoid exposing the main stock to many freeze/thaw cycles which could damage the DNA too. So there is about 50μl fragment volume left on the main IDT tubes that we have received.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+<p>Resulting concentration:</p>
+<table width="587">
+<tbody>
+<tr>
+<td width="148">
+<p>Concentration</p>
+</td>
+<td width="214">
+<p>Aliquoted tube (50&mu;l)</p>
+</td>
+<td width="224">
+<p>IDT Tube (~50&mu;l)</p>
+</td>
+</tr>
+<tr>
+<td width="148">
+<p>NSP4-Tyrosinase</p>
+<p>&nbsp;(1022 bp)</p>
+</td>
+<td width="214">
+<p>10.84 ng/&mu;l = 0.017 pmol</p>
+</td>
+<td width="224">
+<p>8.341 ng/&mu;l = 0.013 pmol</p>
+</td>
+</tr>
+<tr>
+<td width="148">
+<p>OPHT (1495 bp)</p>
+</td>
+<td width="214">
+<p>8.82 ng/&mu;l = 0.0095 pmol</p>
+</td>
+<td width="224">
+<p>7.147 ng/&mu;l = 0.0077 pmol</p>
+</td>
+</tr>
+<tr>
+<td width="148">
+<p>OPHTII (2211 bp)</p>
+</td>
+<td width="214">
+<p>7.65 ng/&mu;l = 0.0055 pmol</p>
+</td>
+<td width="224">
+<p>6.968 ng/&mu;l = 0.0075 pmol</p>
+</td>
+</tr>
+<tr>
+<td width="148">
+<p>OPHTIII (1507 bp)</p>
+</td>
+<td width="214">
+<p>5.68 ng/&mu;l = 0.0060 pmol</p>
+</td>
+<td width="224">
+<p>23.29 ng/&mu;l = 0.02501 pmol</p>
+</td>
+</tr>
+<tr>
+<td width="148">
+<p>OPHM (1782 bp)</p>
+</td>
+<td width="214">
+<p>3.85 ng/&mu;l = 0.00349 pmol</p>
+</td>
+<td width="224">
+<p>5.164 ng/&mu;l = 0.0047 pmol</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p><strong>&nbsp;</strong>Pmol calculations through <a href="https://nebiocalculator.neb.com/#!/dsdnaamt">https://nebiocalculator.neb.com/#!/dsdnaamt</a>.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0614-->
+<!--Modal0615-->
+  <div class="modal" id="modal0615">
+    <div class="modal-content">
+      <div class="modal-header">June 15, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Restriction digestion of pBBR1MCS2</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Restriction digestion reaction: (pBBR1MCS2 concentration:75.5ng/&mu;l)</p>
+<table width="331">
+<tbody>
+<tr>
+<td width="173">
+<p>pBBR1MCS2(2.5mg)</p>
+</td>
+<td width="158">
+<p>2500/75.5&asymp;33ml</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>EcoRI</p>
+</td>
+<td width="158">
+<p>3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>BamHI</p>
+</td>
+<td width="158">
+<p>3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>Cut smart</p>
+</td>
+<td width="158">
+<p>5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>dd H2O</p>
+</td>
+<td width="158">
+<p>6&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>Total</p>
+</td>
+<td width="158">
+<p>50&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Put in thermocycler for 1h. Settings are as following: 37℃ for 1h, 4℃ for infinite</p>
+<p>Gel running: Loading samples (50&mu;l+10 loading dye). Put original plasmid to compare with digested one. Run at 120V for 20 min.</p>
+<p>Gel purification: After excise bands, we did gel purification using protocol from Illustra GFX PCR DNA &amp; Gel Band Purification Kit Gel. Final elution volume is 40&mu;l. Then using Nanodrop to check its concentration which is 7.278ng/&mu;l. It is not a good concentration result. So we repeated the experiment again.</p>
+<p>Repeat restriction digestion: Prepare 4 tubes, 50&mu;l each, total 200&mu;l. The reaction is the same as previous. Overnight digestion at 37℃.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/2/29/T--UM_Macau--Lablog_15_Jun.jpg" style="width:80%; margin-bottom:2%;">
+              <br>
+              <p>Both are the digested products, we just separated them into two wells like usual because one well might overflow and we would lose DNA. Both of them pBBR1MCS2 are at the expected size.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0615-->
+<!--Modal0616-->
+  <div class="modal" id="modal0616">
+    <div class="modal-content">
+      <div class="modal-header">June 16, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat gel purification</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Gel running: This time we put all digested products into one big well. (200&mu;l sample+40&mu;l loading dye). Order is 1kb ladder, sample, and 1 kb ladder. Then Still run at 120V for 20 min.</p>
+<p>&nbsp;</p>
+<p>Gel purification: After excising the gel and following the same protocol, we eluted it with 20&mu;l elution buffer.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/f/fb/T--UM_Macau--Lablog_16_Jun.jpg" style="width: 100%; margin-bottom: 2%;">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0616-->
+<!--Modal0618-->
+  <div class="modal" id="modal0618">
+    <div class="modal-content">
+      <div class="modal-header">June 18, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBBR1MCS2+OPHTIII construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Gibson assembly:</p>
+<table width="625">
+<tbody>
+<tr>
+<td width="313">
+<p>Insert (OPHT III)</p>
+</td>
+<td width="313">
+<p>Vector (pBBR1MCS2)</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Length: 1507 bp</p>
+</td>
+<td width="313">
+<p>Length: 5148 bp</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Concentration: 23.29 ng/&mu;l</p>
+</td>
+<td width="313">
+<p>Concentration: 52.3 ng/&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Volume used for ligation: 3.6 &mu;l</p>
+</td>
+<td width="313">
+<p>Volume used for ligation: 1.91 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Mass: 83.5 ng</p>
+</td>
+<td width="313">
+<p>Mass: 98.4 ng</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Mole: 25.01fmol /0.025 pmol</p>
+<p>(by NEBioCalculator)</p>
+</td>
+<td width="313">
+<p>Mole: 16.44 fmol / 0.016 pmol</p>
+<p>(by NEBioCalculator)</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Volume used (Vector : Insert = 1 : 3)</p>
+<p>Gibson assembly reaction:</p>
+<table width="378">
+<tbody>
+<tr>
+<td width="313">
+<p>Insert (OPHT III)</p>
+</td>
+<td width="65">
+<p>3.6&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Vector (pBBR1MCS2)</p>
+</td>
+<td width="65">
+<p>1.91&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="65">
+<p>10&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Deionized H2O</p>
+</td>
+<td width="65">
+<p>4.49&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="313">
+<p>Total</p>
+</td>
+<td width="65">
+<p>20 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Incubate at 50℃ for 1h.</p>
+<p>Transformation of bacteria: The only changes that we usually make are the following:</p>
+<ol>
+<li>Incubation times for both thawing and incubating with DNA. Varies from 25-30 mins</li>
+<li>The amount of LB broth to put for the outgrowth step varies from 500-900 ul (This case Hugh used 500 ul)</li>
+<li>Incubation time for the outgrowth step varies from 45 mins - 1 hr (This time we used 1 hr)</li>
+<li>Centrifuge 3000 x g for 2 minutes after outgrowth step to pellet the bacteria then remove some supernatant to leave 250 ul LB solution then resuspend the pellet with the leftover 250 ul LB solution then plate all the bacteria on the plate.</li>
+<li>Dry the plates for 10 minutes before inverting and putting into the 37 C incubator</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0618-->
+<!--Modal0619-->
+  <div class="modal" id="modal0619">
+    <div class="modal-content">
+      <div class="modal-header">June 19, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBBR1MCS2+OPHTIII construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Colony picking and incubation</p>
+<ol>
+<li>Picked 6 colonies from the plate</li>
+<li>Put 8 mL LB broth total and added 8 uL Ampicillin</li>
+<li>Put in 37 C shaking incubator overnight for 12-16 hrs</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0619-->
+<!--Modal0620-->
+  <div class="modal" id="modal0620">
+    <div class="modal-content">
+      <div class="modal-header">June 20, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBBR1MCS2+OPHTIII construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Plasmid extraction: using QIAprep Spin Miniprep Kit.</p>
+<p>6 tubes containing 8ml transformed bacteria cultured with LB broth. Pellet 6ml bacterial overnight culture from each tube. Add 250&mu;l buffer D1, then 250&mu;l buffer D2, 350&mu;l buffer N3. Centrifuge at 13000 rpm for 10 min. Transform the 800&mu;l supernatant into spin column. Centrifuge at 13000 rpm for 60s and discard flow through. Add 500&mu;l buffer PB with ethanol. Then centrifuge again at 13000 rpm for 60s and discard flow through. Add 750&mu;l buffer PE, centrifuge at 13000 rpm for 60s and repeat twice. Put the spin columns into new tubes and add 35&mu;l buffer EB. Stand for 5min then centrifuge again. Using Nanodrop to measure their concentration. </p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0620-->
+<!--Modal0626-->
+  <div class="modal" id="modal0626">
+    <div class="modal-content">
+      <div class="modal-header">June 26, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBBR1MCS2+OPHTII Construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+              <p>Gibson assembly</p>
+              <p>Transformation of bacteria</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Failed: subsequent days were used as diagnostic and review</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0626-->
+<!--Modal0627-->
+  <div class="modal" id="modal0627">
+    <div class="modal-content">
+      <div class="modal-header">June 27, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBBR1MCS2+OPHTII Construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+              <p>Colony picking and overnight culture</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0627-->
+<!--Modal0628-->
+  <div class="modal" id="modal0628">
+    <div class="modal-content">
+      <div class="modal-header">June 28, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat pBBR1MCS2+OPHTII Construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+              <p>Gibson assembly</p>
+              <p>Transformation of bacteria</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0628-->
+<!--Modal0629-->
+  <div class="modal" id="modal0629">
+    <div class="modal-content">
+      <div class="modal-header">June 29, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat pBBR1MCS2+OPHTII Construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Plasmid Extraction of pBBR1MCS2: using Midi Kit</p>
+<p>Transfer the overnight culture into four 50ml tubes.</p>
+<p>Nanodrop result is 54.42 ng/ul</p>
+<p>&nbsp;</p>
+<p>Restriction digestion</p>
+<p>PCR for 2 hours at 37 degrees, infinite hold at 4 degree</p>
+<p>&nbsp;</p>
+<p>Gel run for 25 mins at 130V</p>
+<p>(We didnt save the gel image results), we cut the lower band and did gel purification for the pBBR1MCS2 vector.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0629-->
+<!--Modal0630-->
+  <div class="modal" id="modal0630">
+    <div class="modal-content">
+      <div class="modal-header">June 30, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat pBBR1MCS2+OPHTII Construction, Bacteria Transformation</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Gibson Assembly of pBBR1MCS2 and OPHT2</p>
+<p>Run for 1 hour at 50 degrees, infinite hold at 4 degrees</p>
+<p>&nbsp;</p>
+<p>Transformation:</p>
+<p>We thawed the competent cells for only 12 minutes, we should thaw them for a longer time. Then we spread the bacteria on Kan+ agar plates, culture them for 16 hours.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Transformation: It has no colony on positive control plate but few colonies on OPHT2 plate.</p>
+              <div class="row" style="width: 80%; margin-top: 2%">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/b/ba/T--UM_Macau--Lablog_30_Jun_1.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/1/15/T--UM_Macau--Lablog_30_Jun_2.jpg">
+                </div>
+              </div>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0630-->
+<!--July Calendar-->
 <div id="July" class="vertabcontent">
    <div class="wrapper">
@@ Line 869: / Line 2,978: @@
      <div class="days">
        <div><button disabled class="prev_date">30</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">1</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0701">1</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">2</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0702">2</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">3</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0703">3</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">4</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0704">4</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">5</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0705">5</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">6</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0706">6</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">7</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0707">7</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">8</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0708">8</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">9</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0709">9</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">10</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0710">10</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">11</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0711">11</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">12</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0712">12</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">13</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0713">13</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">14</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0714">14</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">15</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0715">15</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">16</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0716">16</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">17</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0717">17</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">18</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0718">18</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">19</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0719">19</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">20</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0720">20</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">21</button></div>
+       <div class="July"><button class="modal-open" data-modal="modal0721">21</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">22</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0722">22</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">23</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0723">23</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">24</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0724">24</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">25</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0725">25</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">26</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0726">26</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">27</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0727">27</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal1">28</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0728">28</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal2">29</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0729">29</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">30</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0730">30</button></div>
-       <div class="July"><button class="modal-open" data-modal="modal3">31</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0731">31</button></div>
      </div>
    </div>
    </div>
 </div>
+<!--End of July Calendar-->
+<!--July Modal-->
+  <div class="modal" id="modal0701">
+    <div class="modal-content">
+      <div class="modal-header">July 1, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat pBBR1MCS2+OPHTII Construction, Start OPHM construction and Transformation of OPHM</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Colony picking and bacteria incubation: Picked 4 colonies from OPHTII agar plate for overnight LB broth culture LB broth volume 40ml + Kanamycin volume 0.04ml</p>
+<p>&nbsp;</p>
+<p>Gibson assembly for pBBR1MCS2+OPHM construction:</p>
+<p>(insert) OPHM concentration = 5.164ng/ul = 0.0047pmol</p>
+<p>(vector) digested pBBR1MCS2 = 12.60ng/ul = 0.004pmol</p>
+<p>&nbsp;</p>
+<p>60ng of vector = 60/12.60 = 4.76ul</p>
+<p>4.76ul x 0.004pmol = 0.019 pmol</p>
+<p>vector:insert=1:2</p>
+<p>Insert = 0.019pmol x 2 =0.038pmol</p>
+<p>Volume for insert = 0.038/0.0047 = 8.09ul</p>
+<p>&nbsp;</p>
+<ol start="2">
+<li>pBBR1MCS2+OPHM (vector:insert=1:2)</li>
+</ol>
+<table width="379">
+<tbody>
+<tr>
+<td width="279">
+<p>Insert (OPHM)</p>
+</td>
+<td width="100">
+<p>8.09 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>60ng Vector (pBBR1MCS2)</p>
+</td>
+<td width="100">
+<p>4.76 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="100">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Deionized H2O</p>
+</td>
+<td width="100">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Total</p>
+</td>
+<td width="100">
+<p>22.85 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<ol start="2">
+<li>Positive control</li>
+</ol>
+<table width="381">
+<tbody>
+<tr>
+<td width="279">
+<p>Insert</p>
+</td>
+<td width="102">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Positive control</p>
+</td>
+<td width="102">
+<p>10ul</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="102">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Deionized H2O</p>
+</td>
+<td width="102">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Total</p>
+</td>
+<td width="102">
+<p>20 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<ol start="3">
+<li>Negative control</li>
+</ol>
+<table width="381">
+<tbody>
+<tr>
+<td width="279">
+<p>Insert</p>
+</td>
+<td width="102">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Vector (pBBR1MCS2)</p>
+</td>
+<td width="102">
+<p>10ul</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="102">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Deionized H2O</p>
+</td>
+<td width="102">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Total</p>
+</td>
+<td width="102">
+<p>20 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Incubate at 50℃ for 1h.</p>
+<p>&nbsp;</p>
+<p>Transformation:</p>
+<p>a. Heat shock at 42℃ for 45 secs using water bath.</p>
+<p>b. Incubate at 37℃ for 60 mins, then plated them into three plates and cultured them for 16 hours.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Transformation: The positive control plate has no colony due to our mistake of plating the bacteria to Kan+ agar plate, it should be plated on Amp+ agar plate.The negative control plate has many colonies which should not happen, we suspect that the plasmid is self-ligated although we transformed with vector without insert. The OPHM agar plate has man colonies which is the expected results.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0702">
+    <div class="modal-content">
+      <div class="modal-header">July 2, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat pBBR1MCS2+OPHTII Construction  Plasmid Extraction of pBBR1MCS2-OPHT2 </p>
+            <p>Restriction digestion for plasmid identity checking</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Plasmid Extraction: We eluted with 40ul elution buffer for x3 times.</p>
+<p>&nbsp;</p>
+<p>Restriction digestion reaction:</p>
+<table width="524">
+<tbody>
+<tr>
+<td width="113">
+<p>&nbsp;</p>
+</td>
+<td width="103">
+<p>C1</p>
+</td>
+<td width="103">
+<p>C2</p>
+</td>
+<td width="103">
+<p>C3</p>
+</td>
+<td width="103">
+<p>C4</p>
+</td>
+</tr>
+<tr>
+<td width="113">
+<p>500ng DNA</p>
+</td>
+<td width="103">
+<p>500/82.28=6.1ul</p>
+</td>
+<td width="103">
+<p>500/87.78=5.7ul</p>
+</td>
+<td width="103">
+<p>500/85.39=5.9ul</p>
+</td>
+<td width="103">
+<p>500/88.74=5.6ul</p>
+</td>
+</tr>
+<tr>
+<td width="113">
+<p>EcoRI</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="113">
+<p>BamHI</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+<td width="103">
+<p>0.5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="113">
+<p>Cut smart</p>
+</td>
+<td width="103">
+<p>1&mu;l</p>
+</td>
+<td width="103">
+<p>1&mu;l</p>
+</td>
+<td width="103">
+<p>1&mu;l</p>
+</td>
+<td width="103">
+<p>1&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="113">
+<p>dd H2O</p>
+</td>
+<td width="103">
+<p>1.9&mu;l</p>
+</td>
+<td width="103">
+<p>2.3&mu;l</p>
+</td>
+<td width="103">
+<p>2.1&mu;l</p>
+</td>
+<td width="103">
+<p>2.4&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="113">
+<p>Total</p>
+</td>
+<td width="103">
+<p>10&mu;l</p>
+</td>
+<td width="103">
+<p>10&mu;l</p>
+</td>
+<td width="103">
+<p>10&mu;l</p>
+</td>
+<td width="103">
+<p>10&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Put in thermocycler for 2hrs. Settings are as following: 37℃ for 2hrs, 4℃ for infinite hold.</p>
+<p>Gel running: Load 4 samples (10&mu;l+2 loading dye) and 1kb ladder (2ul) into wells. Put original plasmid to compare with digested one. Run at 120V for 20 min.</p>
+<p>&nbsp;</p>
+<p>Restriction digestion reaction: (pBBR1MCS2 concentration:75.5ng/&mu;l)</p>
+<table width="331">
+<tbody>
+<tr>
+<td width="173">
+<p>pBBR1MCS2(2ug)</p>
+</td>
+<td width="158">
+<p>40ul</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>EcoRI</p>
+</td>
+<td width="158">
+<p>1&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>BamHI</p>
+</td>
+<td width="158">
+<p>1&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>Cut smart</p>
+</td>
+<td width="158">
+<p>5&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>dd H2O</p>
+</td>
+<td width="158">
+<p>3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="173">
+<p>Total</p>
+</td>
+<td width="158">
+<p>50&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>We made two tubes of the reaction above. Put in thermocycler for 14hrs. Settings are as following: 37℃ for 14hrs, 4℃ for infinite.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Concentration of OPHT2 plasmid:<p>
+              <p>colony 1(C1)=82.28ng/ul</p>
+              <p>colony 2(C2)=87.78ng/ul</p>
+              <p>colony 3(C3)=85.39ng/ul</p>
+              <p>colony 4(C4)=88.74ng/ul</p>
+              <br>
+              <p>Gel run: The expected results should be two bands shown on the gel as the plasmids should be digested by enzyme at the restriction sites. However, there was only a band for each sample, it might due to the plasmid is undigested or not fully digested. Therefore, we have to reconstruct the plasmid backbone.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0703">
+    <div class="modal-content">
+      <div class="modal-header">July 3, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Repeat pBBR1MCS2+OPHTII Construction</p>
+                <p>pBBR1MCS2+OPHM Construction</p>
+                <p>Prepare LB agar plates</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Gel purification: Run gel. Order is 1kb ladder, pBBR1MCS2, pBBR1MCS2, none, OPHT II, pBBR1MCS2, pBBR1MCS2. Used 20&mu;l elution buffer type 6. Concentration is 25.42ng/&mu;l.</p>
+            <br>
+<p>Gibson assembly reaction:</p>
+<p>OPHT II concentration: 7.645ng/&mu;l = 0.0055pmol</p>
+<p>pBBR1MCS2 concentration: 25.42ng/&mu;l = 0.008pmol</p>
+<p>OPHM concentration: 5.164ng/&mu;l = 0.0047pmol</p>
+<p>1.pBBR1MCS2+OPHTII (insert: vector=3:1)</p>
+<table width="379">
+<tbody>
+<tr>
+<td width="279">
+<p>Insert (OPHT II)</p>
+</td>
+<td width="100">
+<p>8.7 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>100 ng Vector (pBBR1MCS2)</p>
+</td>
+<td width="100">
+<p>3 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="100">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Deionized H2O</p>
+</td>
+<td width="100">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Total</p>
+</td>
+<td width="100">
+<p>21.7 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<ol start="2">
+<li>pBBR1MCS2+OPHM (insert: vector=2:1)</li>
+</ol>
+<table width="379">
+<tbody>
+<tr>
+<td width="279">
+<p>100 ng Insert (OPHM)</p>
+</td>
+<td width="100">
+<p>10.21 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Vector (pBBR1MCS2)</p>
+</td>
+<td width="100">
+<p>3 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="100">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Deionized H2O</p>
+</td>
+<td width="100">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Total</p>
+</td>
+<td width="100">
+<p>23.2 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<ol start="3">
+<li>Negative control</li>
+</ol>
+<table width="381">
+<tbody>
+<tr>
+<td width="279">
+<p>Insert</p>
+</td>
+<td width="102">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Vector (pBBR1MCS2)</p>
+</td>
+<td width="102">
+<p>&nbsp;10ul?</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Gibson Assembly Master Mix (2X)</p>
+</td>
+<td width="102">
+<p>10 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Deionized H2O</p>
+</td>
+<td width="102">
+<p>0 &mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="279">
+<p>Total</p>
+</td>
+<td width="102">
+<p>20 &mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Incubate at 50℃ for 1h.</p>
+<p>&nbsp;</p>
+<p>Transformation of OPHT2, OPHM and negative control:</p>
+<p>a. Heat shock at 42℃ for 45secs using water bath.</p>
+<p>b. Incubate at 37℃ for 45 min then plated them into three plates.</p>
+<p>c. Culture them for 12 hrs.</p>
+<p>&nbsp;</p>
+<p>Prepare LB agar plates: 200ml LB agar= 8g powder+ 200ml MilliQ water.</p>
+<p>The ratio for antibiotics and solution is 1:1000. So 200&mu;l Amp/Kan to each kind of plate.</p>
+<p>We made totally 18 plates with Kan+ and 21 plates with Amp+.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0704">
+    <div class="modal-content">
+      <div class="modal-header">July 4, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Colony picking for OPHT2, OPHM Transformation of OPHT1, OPHT3 and NSP4-Tyrosinase</p>
+                <p>Amplify the plasmid</p>
+                <p>Plasmid Extraction of OPHT2, OPHM</p>
+                <p>Restriction digestion to check plasmid identity</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Colony picking: We picked 6 colonies from each transformed plasmids and incubate them in 5mL LB with antibiotic for 12-14 hrs.</p>
+            <br>
+<p>Transformation: Transform OPHT2(#2),&nbsp; OPHT3(#2), NSP4(#6) into Dh5 strain E.coli and incubate at 37 degrees.</p>
+<br>
+<p>Amplify the plasmid by streaking bacteria glycerol stock and culture in 5mL LB broth with antibiotic.</p>
+<br>
+<p>Plasmid Extraction: Save overnight culture before extraction. Used 30ul elution buffer and eluted 3 times, each time incubated 3 minutes. Concentration of DNA is measured by using Nanodrop.</p>
+<p>&nbsp;</p>
+<p>Restriction digestion reaction:</p>
+<table width="630">
+<tbody>
+<tr>
+<td width="97">
+<p>&nbsp;</p>
+</td>
+<td width="89">
+<p>#1</p>
+</td>
+<td width="89">
+<p>#2</p>
+</td>
+<td width="89">
+<p>#3</p>
+</td>
+<td width="89">
+<p>#4</p>
+</td>
+<td width="89">
+<p>#5</p>
+</td>
+<td width="89">
+<p>#6</p>
+</td>
+</tr>
+<tr>
+<td width="97">
+<p>500ng OPHM</p>
+</td>
+<td width="89">
+<p>500/95.65=5.3ul</p>
+</td>
+<td width="89">
+<p>5.3ul</p>
+</td>
+<td width="89">
+<p>5.3ul</p>
+</td>
+<td width="89">
+<p>4.8ul</p>
+</td>
+<td width="89">
+<p>5.1ul</p>
+</td>
+<td width="89">
+<p>5.5ul</p>
+</td>
+</tr>
+<tr>
+<td width="97">
+<p>EcoRI</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="97">
+<p>BamHI</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+<td width="89">
+<p>0.3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="97">
+<p>Cut smart</p>
+</td>
+<td width="89">
+<p>1&mu;l</p>
+</td>
+<td width="89">
+<p>1&mu;l</p>
+</td>
+<td width="89">
+<p>1&mu;l</p>
+</td>
+<td width="89">
+<p>1&mu;l</p>
+</td>
+<td width="89">
+<p>1&mu;l</p>
+</td>
+<td width="89">
+<p>1&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="97">
+<p>dd H2O</p>
+</td>
+<td width="89">
+<p>3.1&mu;l</p>
+</td>
+<td width="89">
+<p>3.1&mu;l</p>
+</td>
+<td width="89">
+<p>3.1&mu;l</p>
+</td>
+<td width="89">
+<p>3.6&mu;l</p>
+</td>
+<td width="89">
+<p>3.3ul</p>
+</td>
+<td width="89">
+<p>2.9ul</p>
+</td>
+</tr>
+<tr>
+<td width="97">
+<p>Total</p>
+</td>
+<td width="89">
+<p>10&mu;l</p>
+</td>
+<td width="89">
+<p>10&mu;l</p>
+</td>
+<td width="89">
+<p>10&mu;l</p>
+</td>
+<td width="89">
+<p>10&mu;l</p>
+</td>
+<td width="89">
+<p>10&mu;l</p>
+</td>
+<td width="89">
+<p>10&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>&nbsp;</p>
+<table width="630">
+<tbody>
+<tr>
+<td width="90">
+<p>&nbsp;</p>
+</td>
+<td width="90">
+<p>#1</p>
+</td>
+<td width="90">
+<p>#2</p>
+</td>
+<td width="90">
+<p>#3</p>
+</td>
+<td width="90">
+<p>#4</p>
+</td>
+<td width="90">
+<p>#5</p>
+</td>
+<td width="90">
+<p>#6</p>
+</td>
+</tr>
+<tr>
+<td width="90">
+<p>500ng OPHT2</p>
+</td>
+<td width="90">
+<p>4.3ul</p>
+</td>
+<td width="90">
+<p>4.1ul</p>
+</td>
+<td width="90">
+<p>5.2ul</p>
+</td>
+<td width="90">
+<p>4.6ul</p>
+</td>
+<td width="90">
+<p>4.7ul</p>
+</td>
+<td width="90">
+<p>4.9ul</p>
+</td>
+</tr>
+<tr>
+<td width="90">
+<p>EcoRI</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="90">
+<p>BamHI</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+<td width="90">
+<p>0.3&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="90">
+<p>Cut smart</p>
+</td>
+<td width="90">
+<p>1&mu;l</p>
+</td>
+<td width="90">
+<p>1&mu;l</p>
+</td>
+<td width="90">
+<p>1&mu;l</p>
+</td>
+<td width="90">
+<p>1&mu;l</p>
+</td>
+<td width="90">
+<p>1&mu;l</p>
+</td>
+<td width="90">
+<p>1&mu;l</p>
+</td>
+</tr>
+<tr>
+<td width="90">
+<p>dd H2O</p>
+</td>
+<td width="90">
+<p>4.1ul</p>
+</td>
+<td width="90">
+<p>4.3ul</p>
+</td>
+<td width="90">
+<p>3.2ul</p>
+</td>
+<td width="90">
+<p>3.8ul</p>
+</td>
+<td width="90">
+<p>3.7ul</p>
+</td>
+<td width="90">
+<p>3.5ul</p>
+</td>
+</tr>
+<tr>
+<td width="90">
+<p>Total</p>
+</td>
+<td width="90">
+<p>10&mu;l</p>
+</td>
+<td width="90">
+<p>10&mu;l</p>
+</td>
+<td width="90">
+<p>10&mu;l</p>
+</td>
+<td width="90">
+<p>10&mu;l</p>
+</td>
+<td width="90">
+<p>10&mu;l</p>
+</td>
+<td width="90">
+<p>10&mu;l</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Put in thermocycler for 4hrs. Settings are as following: 37℃ for 4hrs, 4℃ for infinite hold.</p>
+<p>&nbsp;</p>
+<p>Run the samples on the agarose gel and observe the DNA bands under UV. The estimated correct band size of plasmid colonies are sent for RNA sequencing.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+             <p>Concentration of DNA:</p>
+<p><u>OPHM</u>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <u>OPHT2</u></p>
+<p>#1=95.65ng/ul&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; #1=117.1ng/ul</p>
+<p>#2=94.81ng/ul&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; #2=124.3ng/ul</p>
+<p>#3=94.47ng/ul&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;#3=97.22ng/ul</p>
+<p>#4=204.6ng/ul&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; #4=109.4ng/ul</p>
+<p>#5=98.68ng/ul&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; #5=108.2ng/ul</p>
+<p>#6=91.51ng/ul&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; #6=102.3ng/ul</p>
+<p>&nbsp;</p>
+<p>Gel run and observation:</p>
+<p>OPHT2 (#2 ans #5)</p>
+<p>OPHM(#2 and #6)</p>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0710">
+    <div class="modal-content">
+      <div class="modal-header">July 10, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+       <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>1.Preparation of 20% Arabinose</p>
+                <p>2.SDS-Dye preparation</p>
+                <p>3.Protein samples preparations</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>1: Prepared 5 mL 20% arabinose</p>
+<p>Mixed 1051.9 mg Arabinose + 5 mL milliQ water</p>
+<br>
+<p>2: 4x SDS sdye preparation</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p><strong>Component name</strong></p>
+</td>
+<td width="116">
+<p><strong>Volume</strong></p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>4 x stacking gel buffer</p>
+</td>
+<td width="116">
+<p>25 mL</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>SDS</p>
+</td>
+<td width="116">
+<p>2 g</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Glycerol</p>
+</td>
+<td width="116">
+<p>20 mL</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Bromophenol blue 2-mercaptoethanol</p>
+</td>
+<td width="116">
+<p>20 mg</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>3: Protein samples preparation</p>
+<p>&nbsp;OPHT, OPHTIIII and NSP4 Pellets</p>
+<p>-Resuspended them all with 75 ul Milli-Q water + 25 ul 4 x SDS Page dye. Stored in -20C fridge</p>
+<p>Stored 1 mL of the supernatant of the induced culture tube for later protein expression test.</p>
+<p>Prepared loading sample for supernatant. 75 ul NSP4 supernatant + 25 ul 4x SDS page dye</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0711">
+    <div class="modal-content">
+      <div class="modal-header">July 11, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+          <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pBBR1MCS2+OPHTII(sequenced) Construction</p>
+                <p>pBBR1MCS2+OPHM(sequenced) Construction</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Transformation:</p>
+              <p>2.5ul OPHTII #5 with 50ul competent cell</p>
+              <p>2ul OPHM #2 with 50ul competent cell</p>
+              <p>Plated at 16:15 and incubated at 37 degree overnight.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0712">
+    <div class="modal-content">
+      <div class="modal-header">July 12, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>SDS-Page gel running and Coomassie blue staining</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Gel percentage used 12.5%</p>
+<p>Protein samples that were ran:</p>
+<p>NSP4 = 37.17 kDA</p>
+<p>OPHTI = 52.18 kDA</p>
+<p>OPHTIII = 52.83 kDA</p>
+<p>&nbsp;</p>
+<p>10x Prepared running buffer:</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p><strong>Component name</strong></p>
+</td>
+<td width="116">
+<p><strong>Amount</strong></p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Tris</p>
+</td>
+<td width="116">
+<p>30g</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Glycine</p>
+</td>
+<td width="116">
+<p>144g</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>SDS</p>
+</td>
+<td width="116">
+<p>10g</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Gel running arrangement:</p>
+<p>NSP4 supernatant -&gt; NSP4 pellet -&gt; OPHT3 -&gt; OPHT -&gt; Ladder -&gt; NSP4 supernatant -&gt; NSP4 pellet -&gt; OPHT3 -&gt; OPHT -&gt; Ladder</p>
+<p>Running time: 60V for 1hr then 120 V for 1 hr</p>
+<p>&nbsp;</p>
+<p>Coomasie blue staining:</p>
+<ol>
+<li>Washed the SDS away from the gel</li>
+<li>Soaked the gel with the gel stain for 1 hr</li>
+<li>Wash the excess gel stain with dH2O.</li>
+<li>Put water after 3 times washing, boil for 5 minutes in the microwave then let it shake overnight. Start time 10:27 pm</li>
+<li>Remove the water and take picture</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0716">
+    <div class="modal-content">
+      <div class="modal-header">July 16, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Make glycerol stock of pBBR1MCS2+OPHTII(sequenced) and pBBR1MCS2+OPHM (sequenced)</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>200ul 5% glycerol + 800ul bacteria culture</p>
+            <br>
+            <p>Separate 10 tubes SDS dye</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0719">
+    <div class="modal-content">
+      <div class="modal-header">July 19, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Test adhesion system in three cell lines</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Transformation:&nbsp;</p>
+<p>TOP10&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHT#1(1ul)&nbsp;&nbsp;&nbsp;&nbsp; OPHT3 #3(1ul)&nbsp;&nbsp;&nbsp;&nbsp; 07/05 vector (3.5ul)</p>
+<p>BL-21&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHT#1(1ul)&nbsp;&nbsp;&nbsp;&nbsp; OPHT3 #3(1ul)&nbsp;&nbsp;&nbsp;&nbsp; 07/05 vector (3.5ul)</p>
+<p>MG1655&nbsp; &nbsp; &nbsp; &nbsp; OPHT#1 (1ul)&nbsp;&nbsp;&nbsp;&nbsp; OPHT3 #3(1ul)&nbsp;&nbsp;&nbsp;&nbsp; 07/05 vector (3.5ul)</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0720">
+    <div class="modal-content">
+      <div class="modal-header">July 20, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Test adhesion system in three cell lines</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Pick colony: three colonies per plate for TOP10 and BL-21 with OPHT, OPHT3 and vector, totally 18 tubes, culture at 37 degree shaker overnight</p>
+<p>Transformation:</p>
+<p>TOP10&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHT2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHM</p>
+<p>BL-21&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHT2 &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OPHM</p>
+<p>MG1655&nbsp; &nbsp; &nbsp; &nbsp;OPHT2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHM</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>No colony grew on all MG1655 plates. Repeat the plating with all the remainder MG1655 culture.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal0721">
+    <div class="modal-content">
+      <div class="modal-header">July 21, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Test adhesion system in three cell lines</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Pick colony: three colonies per plate for TOP10 and BL-21 with OPHT2 and OPHM, totally 12 tubes, culture at 37 degree shaker overnight</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Still no colony grew on all MG1655 plates. MG1655 was not suitable for our expression system.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of July Modals-->
+<!--August Calendar-->
 <div id="August" class="vertabcontent">
    <div class="wrapper">
@@ Line 929: / Line 4,567: @@
        <div><button disabled class="prev_date">30</button></div>
        <div><button disabled class="prev_date">31</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">1</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0801">1</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0802">2</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">3</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0803">3</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">4</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0804">4</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0805">5</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0806">6</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0807">7</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal20808">8</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">9</button></div>
+       <div class="August"><button class="modal-open" data-modal="modal0809">9</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">10</button></div>
+       <div class="August"><button class="modal-open" data-modal="modal0810">10</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal2">11</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0811">11</button></div>
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+       <div class="August"><button class="modal-open" data-modal="modal0812">12</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">13</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0813">13</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0814">14</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0815">15</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0816">16</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0817">17</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0818">18</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0819">19</button></div>
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+       <div><button disabled class="modal-open" data-modal="modal0820">20</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">21</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0821">21</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">22</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0822">22</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal2">23</button></div>
+       <div class="August"><button class="modal-open" data-modal="modal0823">23</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">24</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0824">24</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">25</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0825">25</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal2">26</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0826">26</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">27</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0827">27</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal1">28</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0828">28</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal2">29</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0829">29</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">30</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0830">30</button></div>
-       <div class="August"><button class="modal-open" data-modal="modal3">31</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0831">31</button></div>
      </div>
    </div>
    </div>
 </div>
+<!--End of August Calendar-->
+<!--Modal0809-->
+  <div class="modal" id="modal0809">
+    <div class="modal-content">
+      <div class="modal-header">August 9, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Renee, Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>pet11a+OPHT/OPHT2/OPHT3/OPHM/OPHA1/OPHA2 Construction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Gel making</li>
+<li>Gel running: 50ul PCR product + 10ul loading dye, load 30ul per well for pet11a+OPHT/OPHT2/OPHT3/OPHM/OPHA1/OPHA2; 3x50ul PCR product + 30ul loading dye, load 90ul per well for pet11a vector. Run at 120V 15mins</li>
+</ol>
+<p>Loading order: Gel1-ladder/OPHT1/OPHT1/OPHT1/OPHT2/OPHT2</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Gel2-blank/ladder/OPHT3/OPHT3/OPHM/OPHM</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Gel3-ladder/OPHA1/OPHA1/blank/OPHA2/OPHA2</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Gel4-ladder/vector/vector</p>
+<p>&nbsp;</p>
+<p>Gel cut weight: OPHT1&nbsp;&nbsp;&nbsp;&nbsp; 0.36g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Buffer type 3: 360ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHT2&nbsp;&nbsp;&nbsp;&nbsp; 0.27g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 270ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OPHT3&nbsp;&nbsp;&nbsp;&nbsp; 0.31g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 310ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OPHM&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 0.31g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 310ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHA1&nbsp;&nbsp;&nbsp;&nbsp; 0.16g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 160ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Vector1&nbsp;&nbsp;&nbsp; 0.57g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 570ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Vector2&nbsp;&nbsp; 0.43g&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 430ul</p>
+<p>&nbsp;</p>
+<p>Concentration: OPHT1&nbsp;&nbsp; 41ng/ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHT2&nbsp;&nbsp; 38.36ng/ul&nbsp;</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OPHT3&nbsp;&nbsp; 18.95ng/ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; OPHM&nbsp;&nbsp; 21.67ng/ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OPHA1&nbsp; 10.02ng/ul</p>
+<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Vector1+ Vector2&nbsp; 54.12ng/ul</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>No band for OPHA2</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0809-->
+<!--Modal0810-->
+  <div class="modal" id="modal0810">
+    <div class="modal-content">
+      <div class="modal-header">August 10, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Repeat pet11a-OPHA2 Construction</p>
+              <p>Continue pet11a-OPHT/OPHT2/OPHT3/OPHM/OPHA1 Construction</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>1. OPHA2 PCR followed by running gel</p>
+<p>2. Gibson assembly:</p>
+<table width="394">
+<tbody>
+<tr>
+<td width="186">
+<p>OPHT1</p>
+</td>
+<td width="208">
+<p>1.91ul</p>
+</td>
+</tr>
+<tr>
+<td width="186">
+<p>pet11a vector (100ng)</p>
+</td>
+<td width="208">
+<p>1.85ul</p>
+</td>
+</tr>
+<tr>
+<td width="186">
+<p>H2O</p>
+</td>
+<td width="208">
+<p>6.24ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<table width="398">
+<tbody>
+<tr>
+<td width="187">
+<p>OPHT2</p>
+</td>
+<td width="211">
+<p>3ul</p>
+</td>
+</tr>
+<tr>
+<td width="187">
+<p>pet11a vector (100ng)</p>
+</td>
+<td width="211">
+<p>1.85ul</p>
+</td>
+</tr>
+<tr>
+<td width="187">
+<p>H2O</p>
+</td>
+<td width="211">
+<p>5.15ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<table width="397">
+<tbody>
+<tr>
+<td width="185">
+<p>OPHT3</p>
+</td>
+<td width="212">
+<p>4.2ul</p>
+</td>
+</tr>
+<tr>
+<td width="185">
+<p>pet11a vector (100ng)</p>
+</td>
+<td width="212">
+<p>1.85ul</p>
+</td>
+</tr>
+<tr>
+<td width="185">
+<p>H2O</p>
+</td>
+<td width="212">
+<p>3.95ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<table width="397">
+<tbody>
+<tr>
+<td width="187">
+<p>OPHM</p>
+</td>
+<td width="210">
+<p>4.2ul</p>
+</td>
+</tr>
+<tr>
+<td width="187">
+<p>pet11a vector (100ng)</p>
+</td>
+<td width="210">
+<p>1.85ul</p>
+</td>
+</tr>
+<tr>
+<td width="187">
+<p>H2O</p>
+</td>
+<td width="210">
+<p>3.95ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<table width="396">
+<tbody>
+<tr>
+<td width="188">
+<p>OPHA1</p>
+</td>
+<td width="208">
+<p>7.63ul</p>
+</td>
+</tr>
+<tr>
+<td width="188">
+<p>pet11a vector (100ng)</p>
+</td>
+<td width="208">
+<p>1.85ul</p>
+</td>
+</tr>
+<tr>
+<td width="188">
+<p>H2O</p>
+</td>
+<td width="208">
+<p>0.52ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>3. Transformation: 2ul DNA+50ul competent cell Stbl3 Transformed OPHT1, OPHT2, OPHT3, OPHM, OPHA1 and positive ctrl; plated 300ul transformed bacteria on agar plates</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Still no band showed on the gel for OPHA2</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0810-->
+<!--Modal0812-->
+  <div class="modal" id="modal0812">
+    <div class="modal-content">
+      <div class="modal-header">August 12, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Plasmid Extraction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+              <p>Used 30ul elution buffer for elution</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+<p>DNA Concentration (ng/ul):</p>
+<p>&nbsp;</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>OPHT #1</p>
+</td>
+<td width="116">
+<p>109.3</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT #2</p>
+</td>
+<td width="116">
+<p>156.8</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT #3</p>
+</td>
+<td width="116">
+<p>116.4</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT2 #1</p>
+</td>
+<td width="116">
+<p>124.6</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT2 #2</p>
+</td>
+<td width="116">
+<p>116.5</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT2 #3</p>
+</td>
+<td width="116">
+<p>147.4</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT3 #1</p>
+</td>
+<td width="116">
+<p>107.9</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT3 #2</p>
+</td>
+<td width="116">
+<p>105.8</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHT3 #3</p>
+</td>
+<td width="116">
+<p>140.9</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHM #1</p>
+</td>
+<td width="116">
+<p>148.0</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHM #2</p>
+</td>
+<td width="116">
+<p>94.47</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHM #3</p>
+</td>
+<td width="116">
+<p>118.0</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHA #1</p>
+</td>
+<td width="116">
+<p>126.6</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHA #2</p>
+</td>
+<td width="116">
+<p>83.39</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>OPHA #3</p>
+</td>
+<td width="116">
+<p>114.0</p>
+</td>
+</tr>
+</tbody>
+</table>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0812-->
+<!--Modal0823-->
+  <div class="modal" id="modal0823">
+    <div class="modal-content">
+      <div class="modal-header">August 23, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Tyrosinase Activity Assay</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Sample 1: Induced NSP4 pellet Sample 2: Induced NSP4 supernatant</p>
+<p>Sample 3: Uninduced NSP4 pellet</p>
+<p>Sample 4: Uninduced NSP4 supernatant</p>
+<p>&nbsp;</p>
+<p>Homogenize cells with 500ul ice-cold Tyrosinase Assay buffer to perform lysis and keep on ice for 10 minutes followed by centrifugation at 10000xg for 15 minutes at 4 degree. Collect the supernatant and estimate protein concentration by using BCA protein assay kit (protein concentration should range between 1 to 2.5 ug/ul)</p>
+<p>&nbsp;</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0823-->
+<!--September Calendar-->
 <div id="September" class="vertabcontent">
    <div class="wrapper">
@@ Line 985: / Line 5,122: @@
      <div class="days">
-       <div class="September"><button class="modal-open" data-modal="modal1">1</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0901">1</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">2</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0902">2</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">3</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0903">3</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">4</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0904">4</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">5</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0905">5</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">6</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0906">6</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">7</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0907">7</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">8</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0908">8</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">9</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0909">9</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">10</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0910">10</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">11</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0911">11</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">12</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0912">12</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">13</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0913">13</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">14</button></div>
+       <div><button disabled class="modal-open" data-modal="modal0914">14</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">15</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0915">15</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">16</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0916">16</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">17</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0917">17</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">18</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0918">18</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">19</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0919">19</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">20</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0920">20</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">21</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0921">21</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">22</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0922">22</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">23</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0923">23</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">24</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0924">24</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">25</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0925">25</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">26</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0926">26</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">27</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0927">27</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal1">28</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0928">28</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal2">29</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0929">29</button></div>
-       <div class="September"><button class="modal-open" data-modal="modal3">30</button></div>
+       <div class="September"><button class="modal-open" data-modal="modal0930">30</button></div>
      </div>
    </div>
    </div>
 </div>
+<!--End of September Calendar-->
+<!--Modal0903-->
+  <div class="modal" id="modal0903">
+    <div class="modal-content">
+      <div class="modal-header">September 3, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Prepare NSP4 for Negative control</p>
+              <p>NSP4 activity test preparation</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Took one tip of cells from glycerol stock, incubate with 5ml LB broth + 5ul Ampicillin at 37 degree shaker at 17:05</li>
+<li>Pull down assay for NSP4 induced supernatant.</li>
+<li>Sonication for NSP4 bacteria culture</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0903-->
+<!--Modal0908-->
+  <div class="modal" id="modal0908">
+    <div class="modal-content">
+      <div class="modal-header">September 8, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Nickel pull-down assay</p>
+                <p>Tyrosinase activity test</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Prepare nickel resin: Pipette 30ul of nickel resin. Wash resin in 1ul H2O two times followed by washing with 1ml binding buffer two times.</li>
+<li>Pull-down assay: Centrifuge the induced BL-21 (NSP4 transformed colony 3) as well as the uninduced one and take the supernatant as pull-down assay sample. Pipette 30ul resuspend resin into supernatant. Incubate and rotate it at 4 degree for&nbsp; 45mins. Spin at 3900rpm for 10mins, remove the supernatant which will be stored just in case, then leave 1ml fraction in&nbsp; resuspend resin and transfer resin to&nbsp; 1.5ml tube. Spin down at 4000g 2min to get resin. Discard supernatant. Wash resin with 750ul binding buffer three times (we washed the resin with Tyrosinase Assay buffer in the third wash). Measure protein concentrationProceed to Tyrosinase Activity Assay.</li>
+<li>Tyrosinase activity test: Homogenize cells with 500ul ice-cold Tyrosinase Assay buffer to perform lysis and keep on ice for 10 minutes followed by centrifugation at 10000xg for 15 minutes at 4 degree. Collect the supernatant and estimate protein concentration by using BCA protein assay kit (protein concentration should range between 1 to 2.5 ug/ul)</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+<table>
+<tbody>
+<tr>
+<td width="215">
+<p>Protein</p>
+</td>
+<td width="162">
+<p>Concentration</p>
+</td>
+</tr>
+<tr>
+<td width="215">
+<p>Uninduced pellet</p>
+</td>
+<td width="162">
+<p>1.67ug/ul</p>
+</td>
+</tr>
+<tr>
+<td width="215">
+<p>Induced pellet</p>
+</td>
+<td width="162">
+<p>5.64 ug/ul</p>
+</td>
+</tr>
+<tr>
+<td width="215">
+<p>Uninduced supernatant</p>
+</td>
+<td width="162">
+<p>-0.19ug/ul</p>
+</td>
+</tr>
+<tr>
+<td width="215">
+<p>Induced supernatant</p>
+</td>
+<td width="162">
+<p>-0.11ug/ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>As the induced supernatant concentration is negative, there is literally no protein in the supernatant, therefore we did not continue proceed the Tyrosinase Activity test.</p>
+<br>
+<div class="row" style="width: 100%; margin-top: 1%;">
+  <div class="col-sm-6">
+    <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/f/f2/T--UM_Macau--Lablog_8_Sep_1.jpg">
+  </div>
+  <div class="col-sm-6">
+    <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/7/77/T--UM_Macau--Lablog_8_Sep_2.jpg">
+  </div>
+</div>
+<div class="row" style="width: 100%; margin-top: 1%;">
+  <div class="col-sm-6">
+    <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/7/71/T--UM_Macau--Lablog_8_Sep_3.jpg">
+  </div>
+  <div class="col-sm-6">
+    <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/b/b1/T--UM_Macau--Lablog_8_Sep_4.jpg">
+  </div>
+</div>
+<div class="row" style="width: 100%; margin-top: 1%;">
+  <div class="col-sm-6">
+    <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/3/31/T--UM_Macau--Lablog_8_Sep_5.jpg">
+  </div>
+  <div class="col-sm-6">
+    <img class="img-fluid" src="">
+  </div>
+</div>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0908-->
+<!--Modal0909-->
+  <div class="modal" id="modal0909">
+    <div class="modal-content">
+      <div class="modal-header">September 9, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Make an overnight culture of BL-21 (NSP4 transformed colony 3) from glycerol stock</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Make a 20mL culture of BL-21 glycerol stock (NSP4 transformed colony 3) with Ampicillin antibody (1:1000 dilution) and start incubation in 37 degree shaking incubator at 6.25pm</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0909-->
+<!--Modal0910-->
+  <div class="modal" id="modal0910">
+    <div class="modal-content">
+      <div class="modal-header">September 10, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>pBAD24 NSP4 #3 transformation with BL-21</p>
+              <p>Induce overnight BL-21 (NSP4 transformed colony 3)</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Transformation: 1.5ul pBAD24 NSP4 #3 + 50ul BL-21 competent cell</li>
+<li>Induction: separate 5ml out of 20ml overnight BL-21 as uninduced BL-21 stock and put into -80 degree, for the rest of 15ml, add 150ul 20% arabinose for induction, then culture overnight in 37 degree shaker</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0910-->
+<!--Modal0911-->
+  <div class="modal" id="modal0911">
+    <div class="modal-content">
+      <div class="modal-header">September 11, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Preparation for NSP4 activity assay</p>
+              <p>Colony picking</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Picked a colony from the transformation yesterday. Prepared a 20 mL culture with amp+ and incubated in 37 C at 6:49 am</li>
+<li>Centrifuge the overnight induced culture of NSP4 Colony #3 BL-21, separate pellet and supernatant, then store in -80degree. Centrifuged 5mL uninduced sample NSP4_plasmid#3 then stored in -80C. Induced the rest 15 mL (OD=1.1227 13-14 hr culture) with 150 ul 20% arabinose at 20:08 and incubated in shaking incubator.</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0911-->
+<!--Modal0912-->
+  <div class="modal" id="modal0912">
+    <div class="modal-content">
+      <div class="modal-header">September 12, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>NSP4 activity assay</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Nickel pull down both induced and uninduced Nsp4 SUPERNATANT (new and old stock), total 4 samples</li>
+<li>Scrapped OPHT2 bacteria (three stocks) from glycerol stock 082719 and cultured in ~5mL volume of Lb broth with amp+ and then put in 37C shaking incubator at 16:45. Glycerol stock for all OPHT2 colonies were thawed completely by accident watch out for the quality of this stock for next scrapping.</li>
+<li>Homogenize 4 samples with tyrosinase assay buffer and test protein concentration with BCA protein assay (30mins incubation)</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0912-->
+<!--Modal0913-->
+  <div class="modal" id="modal0913">
+    <div class="modal-content">
+      <div class="modal-header">September 13, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>OPHT2 induction</p>
+              <p>NSP4 activity assay</p>
+              <p>Material stock preparation</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a> <a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Induced 5mL cultures of pet11a_OPHT2 from yesterday with 1mM IPTG (10 ul of 500mM IPTG stock) and incubated at 37 C shaking incubator at 4:45. OD600 =0.7144</li>
+<li>Tyrosinase activity assay started plate OD monitoring (510nm) at 8:02 am in the plate reader in 37 C temperature for 1 hr. Stored all proteins back in the &ldquo;Protein sample box&rdquo; in -80C</li>
+<li>Made two 500mL LB broth and one 110 mL LB agar for making 10 amp+ plates. Made a stock of 50 mL LB Broth amp+ and put in 4 degree fridge.</li>
+<li>Diluted induced pet11a_OPHT2 #1 from early this morning. 1:1000 dilution ratio by putting 10 ul of that induced culture into 10 mL LB Broth amp+ then stored in shaking incubator 37 C overnight.</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Result of tyrosinase activity test today</p>
+              <img class="img-fluid" style="width: 80%;margin-top:2%;" src="https://static.igem.org/mediawiki/2019/7/73/T--UM_Macau--Lablog_13_Sep.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0913-->
+<!--Modal0915-->
+  <div class="modal" id="modal0915">
+    <div class="modal-content">
+      <div class="modal-header">September 15, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>BL-21 OPHT2 #1 EGFP expression</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a> <a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>eGFP expression is still very high even after ~28 hrs after induction for our bacteria Bl-21 OPHT2 colony #1 </p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" style="width: 80%;margin-top:2%;" src="https://static.igem.org/mediawiki/2019/7/7f/T--UM_Macau--Lablog_15_Sep.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0915-->
+<!--Modal0916-->
+  <div class="modal" id="modal0916">
+    <div class="modal-content">
+      <div class="modal-header">September 16, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Material stock preparation</p>
+              <p>BL-21 OPHT2 #1 EGFP expression</p>
+              <p>Adhesion system reconstruction</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<ol>
+<li>Made 11 Amp+ agar plates by adding 100ul (1:1000) to 100ml agar solution.</li>
+<li>eGFP expression is still very high even after ~40 hrs after induction for our bacteria Bl-21 OPHT2 colony #1</li>
+<li>PCR reaction for the amplification of adhesion system for ligation to pet11a for 1 hr and 26 mins. Started at 15:34</li>
+</ol>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" style="width: 80%;margin-top:2%;" src="https://static.igem.org/mediawiki/2019/6/6f/T--UM_Macau--Lablog_16_Sep.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0916-->
+<!--Modal0917-->
+  <div class="modal" id="modal0917">
+    <div class="modal-content">
+      <div class="modal-header">September 17, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee, Jia Ying</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Adhesion system reconstruction</p>
+              <p>Preparation digested pet11a vector stock</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>1. Incubated a new BL_21 OPHT2 colony #1, 5 mL volume, at 37 C</p>
+<p>2. Run the gel with opht1,2,3 and opha1 samples (each well 30ul, each sample 2 wells) at 120V for 20 mins. Then gel purification.</p>
+<p>3. Gibson assembly to ligate Pet11a vector with Opht1,2,3 and Opha1 inserts. Calculation as below:</p>
+<table style="width: 231.2px;">
+<tbody>
+<tr>
+<td style="width: 49px;">
+<p>Plasmid</p>
+</td>
+<td style="width: 116px;">
+<p>Concentration(ng/ul)</p>
+</td>
+<td style="width: 47.2px;">
+<p>pMol</p>
+</td>
+</tr>
+<tr>
+<td style="width: 49px;">
+<p>pet11a vector</p>
+</td>
+<td style="width: 116px;">
+<p>54.12</p>
+</td>
+<td style="width: 47.2px;">
+<p>0.015</p>
+</td>
+</tr>
+<tr>
+<td style="width: 49px;">
+<p>OPHT1</p>
+</td>
+<td style="width: 116px;">
+<p>6.7</p>
+</td>
+<td style="width: 47.2px;">
+<p>0.0068</p>
+</td>
+</tr>
+<tr>
+<td style="width: 49px;">
+<p>OPHT2</p>
+</td>
+<td style="width: 116px;">
+<p>7.4</p>
+</td>
+<td style="width: 47.2px;">
+<p>0.0051</p>
+</td>
+</tr>
+<tr>
+<td style="width: 49px;">
+<p>OPHT3</p>
+</td>
+<td style="width: 116px;">
+<p>4.5</p>
+</td>
+<td style="width: 47.2px;">
+<p>0.0045</p>
+</td>
+</tr>
+<tr>
+<td style="width: 49px;">
+<p>OPHA1</p>
+</td>
+<td style="width: 116px;">
+<p>17.3</p>
+</td>
+<td style="width: 47.2px;">
+<p>0.018</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>75ng of vector=75/54.12=1.39ul</p>
+<p>1.39ulx0.015pmol=0.02pmol</p>
+<p>vector:insert=1:3</p>
+<p>Insert=0.06pmol</p>
+<table style="width: 348.8px;">
+<tbody>
+<tr>
+<td style="width: 27px;">
+<p>Insert</p>
+</td>
+<td style="width: 47px;">
+<p>Volume of insert(ul)</p>
+</td>
+<td style="width: 58px;">
+<p>Volume of vector(ul)</p>
+</td>
+<td style="width: 54px;">
+<p>Volume of water(ul)</p>
+</td>
+<td style="width: 62px;">
+<p>Volume of Master Mix(ul)</p>
+</td>
+<td style="width: 80.8px;">
+<p>Total volume</p>
+<p>(ul)</p>
+</td>
+</tr>
+<tr>
+<td style="width: 27px;">
+<p>OPHT1</p>
+</td>
+<td style="width: 47px;">
+<p>8.82</p>
+</td>
+<td style="width: 58px;">
+<p>1.39</p>
+</td>
+<td style="width: 54px;">
+<p>-</p>
+</td>
+<td style="width: 62px;">
+<p>10</p>
+</td>
+<td style="width: 80.8px;">
+<p>20.21</p>
+</td>
+</tr>
+<tr>
+<td style="width: 27px;">
+<p>OPHT2</p>
+</td>
+<td style="width: 47px;">
+<p>11.76</p>
+</td>
+<td style="width: 58px;">
+<p>1.39</p>
+</td>
+<td style="width: 54px;">
+<p>-</p>
+</td>
+<td style="width: 62px;">
+<p>13</p>
+</td>
+<td style="width: 80.8px;">
+<p>26.15</p>
+</td>
+</tr>
+<tr>
+<td style="width: 27px;">
+<p>OPHT3</p>
+</td>
+<td style="width: 47px;">
+<p>13.33</p>
+</td>
+<td style="width: 58px;">
+<p>1.39</p>
+</td>
+<td style="width: 54px;">
+<p>-</p>
+</td>
+<td style="width: 62px;">
+<p>14</p>
+</td>
+<td style="width: 80.8px;">
+<p>28.72</p>
+</td>
+</tr>
+<tr>
+<td style="width: 27px;">
+<p>OPHA1</p>
+</td>
+<td style="width: 47px;">
+<p>3.33</p>
+</td>
+<td style="width: 58px;">
+<p>1.39</p>
+</td>
+<td style="width: 54px;">
+<p>5.28</p>
+</td>
+<td style="width: 62px;">
+<p>10</p>
+</td>
+<td style="width: 80.8px;">
+<p>20</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>4.Restriction digestion for pet11a vector. We did it for 100ul, incubate for 14hrs at 37 degree, infinite hold at 4 degree. Calculation as below:</p>
+<p>pET11a concentration: 49.1ng/ul</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>pET11a (3ug)</p>
+</td>
+<td width="116">
+<p>60ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>BamHI</p>
+</td>
+<td width="116">
+<p>2ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>NHeI</p>
+</td>
+<td width="116">
+<p>2ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Cutsmart buffer</p>
+</td>
+<td width="116">
+<p>10ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>ddH2O</p>
+</td>
+<td width="116">
+<p>26ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Total</p>
+</td>
+<td width="116">
+<p>100ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>5. Transformation stock OPHT1, OPHT2, OPHT3, OPHM, and OPHA using BL-21; stock pBBR1MCS2_OPHT2 using BL-21; stock pet11a_OPHT1, OPHT2 using Rosetta. Put into the incubator at 21:00</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Transformation:</p>
+              <div class="row" style="width: 80%; margin-top:1%;">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/c/c4/T--UM_Macau--Lablog_17_Sep_1.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/d/de/T--UM_Macau--Lablog_17_Sep_2.jpg">
+                </div>
+              </div>
+              <div class="row" style="width: 80%; margin-top:1%;">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/2/20/T--UM_Macau--Lablog_17_Sep_3.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/8/8f/T--UM_Macau--Lablog_17_Sep_4.jpg">
+                </div>
+              </div>
+              <div class="row" style="width: 80%; margin-top:1%;">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/a/aa/T--UM_Macau--Lablog_17_Sep_5.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/c/ce/T--UM_Macau--Lablog_17_Sep_6.jpg">
+                </div>
+              </div>
+              <div class="row" style="width: 80%; margin-top:1%;">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/2/27/T--UM_Macau--Lablog_17_Sep_7.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/d/d9/T--UM_Macau--Lablog_17_Sep_8.jpg">
+                </div>
+              </div>
+              <div class="row" style="width: 80%; margin-top:1%;">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/f/fe/T--UM_Macau--Lablog_17_Sep_9.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="">
+                </div>
+              </div>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0917-->
+<!--Modal0918-->
+  <div class="modal" id="modal0918">
+    <div class="modal-content">
+      <div class="modal-header">September 18, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien, Renee, Jia Ying</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Colony picking for transformed bacteria</p>
+              <p>Bacteria transformation </p>
+              <p>Ammonia iron (III) citrate solution preparation</p>
+              <p>Induction of IPTG and Ammonia iron (III) citrate in OPHT2 bacteria</p>
+              <p>Gel run and gel purification</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Colony picking: Picked 1 colony for OPHT1,2,3,M and OPHA1 (BL-21), OPHT1,2 (Rosetta). Picked 3 colonies for pBBR1-Opht2 (BL-21). Incubate the picked colonies with 5ml Amp+ (for pET11a plasmid) or Kan+ (for pBBR1MCS2 plasmid) LB broth, put in 37degree shaking incubator at 4.30pm.</p>
+            <br>
+            <p>Transformation with new constructed pet11a_OPHT, OPHT2, OPHT3 and OPHA using the Stbl3 strain E.coli. Put them into 37 degree incubator at 18:40.</p>
+            <br>
+            <p>Prepared the Ammonia iron (III) citrate solution at concentration:100mM with 26.3g iron(III) powder in 263mL water at 18:00.</p>
+            <br>
+            <p>We added 10ul IPTG and 1ml ammonium iron(iii) citrate into BL-21 OPHT2 at 6pm. We run the gel for digested pET11a vector, the expected band size is 5.6kb. Then we purify the excised gel (band1 weight 0.371g and band2 weight 0.372g) with 30ul elution buffer type 4 three times, each time incubated 5minutes.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Gel run results:</p>
+              <img class="img-fluid" style="width: 50%;"src="https://static.igem.org/mediawiki/2019/6/6e/T--UM_Macau--Lablog_18_Sep.jpg">
+              <br>
+              <p>The concentration for band1 is 2.24ng/ul and band 2 is 2.49ng/ul.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0918-->
+<!--Modal0919-->
+  <div class="modal" id="modal0919">
+    <div class="modal-content">
+      <div class="modal-header">September 19, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Glycerol stock making</p>
+                <p>Colony picking and bacteria overnight culture</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Picked up adhesion system plasmids transformed plates from 37C, parafilmed and put it in 4C. All constructs have colonies. Picked three colonies for each transformed plasmids in Stbl3 with 5ml LB, incubated in 37 shaking incubator at 6.50pm.</p>
+            <br>
+            <p>Collected the overnight culture of picked colonies at 1.30pm [amplified adhesion system-old stock: OPHT1,2,3,M and OPHA1 (BL-21), OPHT1,2 (Rosetta) and pBBR1-Opht2 #1,2,3 (BL-21)]. Made 1ml glycerol stock of these with 500ul 50% glycerol and 500ul bacteria culture, stored in -80 degree fridge’s  Glycerol stock box.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Checked EGFP expression of pbbr1-opht2 with fluorescence microscope:</p>
+              <div class="row" style="width: 100%; margin-top: 1%">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/2/2a/T--UM_Macau--Lablog_19_Sep_1.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/0/03/T--UM_Macau--Lablog_19_Sep_2.jpg">
+                </div>
+              </div>
+              <div class="row" style="width: 100%; margin-top: 1%">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/5/5b/T--UM_Macau--Lablog_19_Sep_3.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/6/6c/T--UM_Macau--Lablog_19_Sep_4.jpg">
+                </div>
+              </div>
+              <div class="row" style="width: 100%; margin-top: 1%">
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/1/1b/T--UM_Macau--Lablog_19_Sep_5.jpg">
+                </div>
+                <div class="col-sm-6">
+                  <img class="img-fluid" src="">
+                </div>
+              </div>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0919-->
+<!--Modal0920-->
+  <div class="modal" id="modal0920">
+    <div class="modal-content">
+      <div class="modal-header">September 20, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Sequencing samples preparation</p>
+                <p>Glycerol stock making</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Collected overnight culture of picked colonies (pet11a-stbl3) and prepared sequencing samples(1ml bacteria, 120ul 106FOR, 30ul 102 FOR), sent for sequencing.</p>
+            <br>
+            <p>Made 1ml glycerol stock with 500ul 50% glycerol stock and 500ul bacteria culture, stored in -80 degree fridge’s glycerol stock box.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0920-->
+<!--Modal0921-->
+  <div class="modal" id="modal0921">
+    <div class="modal-content">
+      <div class="modal-header">September 21, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Protein extraction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Protein extraction for overnight bacteria culture (pet11a adhesion system-BL21, Rosetta) with 500ul lysis buffer, boiled at 98degree for 10minutes. Measured protein concentration after 30minutes BCA incubation. Run western blot at 1.40pm.</p>
+            <br>
+            <p>The order and volume loaded shown as below:</p>
+            <p>Ladder - OPHT1 (Rosetta) - OPHT2(Rosetta) - OPHT1 (BL-21) - OPHT2 (BL-21) - OPHT3 - OPHM - OPHA1</p>
+            <p>(5ul for ladder, 20ul for the rest protein samples)</p>
+            <br>
+            <p>Incubated with antibody and shaking in 4 degrees fridge at 5.20pm</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0921-->
+<!--Modal0922-->
+  <div class="modal" id="modal0922">
+    <div class="modal-content">
+      <div class="modal-header">September 22, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Continue western blot</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Washed the membrane with about 3 times.</p>
+            <br>
+            <p>Incubated 10 mL OPHT2 Colony #1 in LB amp+ at 37 C shaking incubator at 23:30.</p>
+            <p>Magnetization of E.coli incubated in Ferric ammonium citrate(3 days incubation at 37 C) for 3 hrs with magnet. Made a video from the fluorescence microscope</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Western Blot results:</p>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/1/1c/T--UM_Macau--Lablog_22_Sep.jpg" style="width: 60%; margin-top:1%;">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0922-->
+<!--Modal0923-->
+  <div class="modal" id="modal0923">
+    <div class="modal-content">
+      <div class="modal-header">September 23, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Incubation of bacteria culture, adhesion system functional test</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Incubated 15 mL culture of all the new pET11a adhesion system glycerol stock at 37 C shaking incubator at 14:01. Will extract after 12 hrs.</p>
+            <br>
+            <p>Induced the overnight BL-21 with 1mM IPTG (50ul in 10 mL) and put in 37C shaking incubator. Inducing for 3 hrs.</p>
+            <br>
+            <p>Added 2 ml of 100mM ferric ammonium citrate into the 3 hr IPTG induced BL-21 OPHT2 colony #1(Checked egfp signal and it’s positive) and put it in 37C shaking incubator at 17:08. Incubating for 24 hrs.</p>
+            <br>
+            <p>Made 5 mL overnight culture of BL-21 for adhesion system functional test. Stored in 37 C shaking incubator at 20:50 . Used all colony #1 from the old pet11a glycerol stock. Incubating for 12 hrs.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0923-->
+<!--Modal0924-->
+  <div class="modal" id="modal0924">
+    <div class="modal-content">
+      <div class="modal-header">September 24, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Plasmid extraction</p>
+              <p>Induction of adhesion system</p>
+              <p>Magnetization test set up</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a> <a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Extracted plasmids of the 15 mL new pet11a adhesion system and sent samples for sequencing.</p>
+            <br>
+            <p>Stored new pet11a plasmids in -20C in the new construct box.</p>
+            <br>
+            <p>Induced 5 mL overnight for functional test of adhesion system using 1mM IPTG(10 ul 500 mM stock -> 5 mL culture) for 3 hrs in 30C shaking incubator in Prof.Xu’s lab started at 16:30. </p>
+            <br>
+            <p>Started the magnetization test setups at 00:05. Prepared three dishes. Two that was incubated with fe3+ and one negative ctrl. One of the two dishes has been put in the fluorescence microscope and picture is being taken at 30 sec intervals for 6 hrs. </p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0924-->
+<!--Modal0925-->
+  <div class="modal" id="modal0925">
+    <div class="modal-content">
+      <div class="modal-header">September 25, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Incubation of bacteria culture</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Started the incubation of overnight culture of Tyrosinase (500 U/ml) 2 mL + 2 mL of our adhesion system bacteria that was resuspended with 10 uM Phosphate buffer (Which prepared at 092419). Incubated in Professor Xu's lab's shaking incubator temperature 25-27 C, shaking at 75 rpm. </p>
+            <br>
+            <p>Picked up our overnight culture E.coli with Tyrosinase at 19:45 (Total 17 hrs incubation).</p>
+            <br>
+            <p>Started the incubation of our "Activated E.coli with Tyrosinase" at 20:30. Mixed 150 ul of the nanoparticle PFODTBT (30 ppm) with the 4 mL overnight culture. Put the solution in Professor Xu's shaking incubator at 25C 70 rpm for 1 hr.</p>
+            <br>
+            <p>Put the activated bacteria incubated with the nanoparticle in the 4 Degree of PRofessor Xu's lab.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/8/8b/T--UM_Macau--Lablog_25_Sep.jpg" style="width: 80%;">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0925-->
+<!--Modal0926-->
+  <div class="modal" id="modal0926">
+    <div class="modal-content">
+      <div class="modal-header">September 26, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Checking of nanoparticle binding with bacteria</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Centrifuged the nanoparticles incubated bacteria culture and resuspended the pellet with 1mL phosphate buffer. Get 10ul on glass slide and observed under a fluorescence microscope. We have negative ctrl(without tyrosinase), nanoparticles only and 5 bacteria samples.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0926-->
+<!--Modal0927-->
+  <div class="modal" id="modal0927">
+    <div class="modal-content">
+      <div class="modal-header">September 27, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Bacteria culture</p>
+                <p>Restriction digestion</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Prepare 5mL of pet11a adhesion system BL-21 bacteria culture with Amp+, shaking in 37 incubator of Prof. Henry’s lab.</p>
+            <br>
+            <p>Restriction digestion for pBAD4 vector and pBAD4-tyrosine vector with XbaI enzyme. Put in thermocycler 37 degree for 14 hours and 4 degree for infinite hold.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0927-->
+<!--Modal0928-->
+  <div class="modal" id="modal0928">
+    <div class="modal-content">
+      <div class="modal-header">September 28, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>DNA purification</p>
+              <p>Induction of samples</p>
+              <p>Gibson assembly</p>
+              <p>Transformation</p>
+            </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Separated 1ml culture as uninduced sample for each. Induced with 10uL IPTG for each samples except negative ctrl and incubated in Prof’s Xu lab 30 degree shaking incubator at around 11.40pm.</p>
+            <br>
+            <p>Purified DNA of digested pBAD24 and pBAD24_NSP4 from agarose gels. Concentration of pBAD24 is 125.1ng/ul and pBAD24_NSP4 is 61.31ng/ul.</p>
+            <br>
+            <p>Resuspend ftnA fragment with 25ul TE buffer followed by 50degree incubation for 20min. Gibson assembly of ftnA+pBAD24 and ftnA+pBAD24_NSP4.</p>
+            <br>
+            <p>Gibson assembly for pBAD vector+ftnA and pBAD NSP4 +ftnA. Put in thermocycler for 1hr at 50degree. Calculation as below:</p>
+            <p>pBAD NSP4 conc=61.31ng/ul = 0.017pmol</p>
+            <p>pBAD conc=125.1ng/ul = 0.042pmol</p>
+            <p>ftnA conc=20ng/ul = 0.052pmol</p>
+            <br>
+            <p>100ng vector (pBAD NSP4)=1.63ul</p>
+            <p>(vector)1.63 x 0.017=0.028pmol</p>
+            <p>vector:insert=1:3</p>
+            <p>(insert)=0.083pmol</p>
+            <p>ftnA volume=0.083/0.052=1.6ul</p>
+            <table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>Vector</p>
+</td>
+<td width="116">
+<p>1.63ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Insert</p>
+</td>
+<td width="116">
+<p>1.6ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Master mix</p>
+</td>
+<td width="116">
+<p>10ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>water</p>
+</td>
+<td width="116">
+<p>6.77ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>total</p>
+</td>
+<td width="116">
+<p>20ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>100ng vector(pBAD)=0.8ul</p>
+<p>(vector)0.8 x 0.042=0.034pmol</p>
+<p>vector:insert=1:3</p>
+<p>(insert)=0.102pmol</p>
+<p>ftnA volume=0.102/0.052=1.96ul</p>
+<p>&nbsp;</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>Vector</p>
+</td>
+<td width="116">
+<p>0.8ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Insert</p>
+</td>
+<td width="116">
+<p>1.96ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Master mix</p>
+</td>
+<td width="116">
+<p>10ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>water</p>
+</td>
+<td width="116">
+<p>7.29ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>total</p>
+</td>
+<td width="116">
+<p>20ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Incubated our 1 mL bacteria with tyrosinase 500U/mL in Professor Xu&rsquo;s shaking incubator at 25 C 90 rpm for 24 hrs.</p>
+<p>&nbsp;</p>
+<p>Transformation of ftnA+pBAD24 and ftnA+pBAD24_NSP4 into stbl3 strain. Start incubation at 22:17.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0928-->
+<!--Modal0929-->
+  <div class="modal" id="modal0929">
+    <div class="modal-content">
+      <div class="modal-header">September 29, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Colony picking</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Picked three colonies for FtnA+pBAD24 and made 25 mL bacteria solution with amp+ at 17:30. Unfortunately there were no colonies for the pBAD24_NSP4+FtnA. Will repeat this transformation later.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0929-->
+<!--Modal0930-->
+  <div class="modal" id="modal0930">
+    <div class="modal-content">
+      <div class="modal-header">September 30, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Plasmid extraction</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Plasmid extraction for three overnight culture of pbad24-ftna. Used 30ul EB, repeated 3times and each time incubated 5 mins. Extracted concentration listed in "Results"</p>
+            <br>
+            <p>Incubated 100 ul of activated adhesion system + bacteria from 092819 with 10 ul (1 mg/mL concentration) of 1.0%w/v FP-00552-2 Yellow fluoresscent particle, 0.04-0.09 um size in 25C and mixing at 300 rpm for 1 hr. Started at 23:40 (Samples were covered with aluminum foil to prevent exposure to light).</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+<p>Concentration of extracted plasmid:</p>
+<p>pBAD24-ftnA #1: 169.3ng/ul</p>
+<p>pBAD24-ftnA #2: 200.9ng/ul</p>
+<p>pBAD24-ftnA #3: 137.8ng/ul</p>
+<img class="img-fluid" style="width: 80%; margin-top: 1%"; src="https://static.igem.org/mediawiki/2019/1/10/T--UM_Macau--Lablog_30_Sep.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal0930-->
+<!--October Calendar-->
 <div id="October" class="vertabcontent">
    <div class="wrapper">
@@ Line 1,042: / Line 6,904: @@
        <div><button disabled class="prev_date">29</button></div>
        <div><button disabled class="prev_date">30</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">1</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1001">1</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">2</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1002">2</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">3</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1003">3</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">4</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1004">4</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">5</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1005">5</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">6</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1006">6</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">7</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1007">7</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">8</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1008">8</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">9</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1009">9</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">10</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1010">10</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">11</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1011">11</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">12</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1012">12</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">13</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1013">13</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">14</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1014">14</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">15</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1015">15</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">16</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1016">16</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">17</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1017">17</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">18</button></div>
+       <div class="October"><button class="modal-open" data-modal="modal1018">18</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">19</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1019">19</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">20</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1020">20</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">21</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1021">21</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">22</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1022">22</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">23</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1023">23</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">24</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1024">24</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">25</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1025">25</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">26</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1026">26</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">27</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1027">27</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal1">28</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1028">28</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal2">29</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1029">29</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">30</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1030">30</button></div>
-       <div class="October"><button class="modal-open" data-modal="modal3">31</button></div>
+       <div><button disabled class="modal-open" data-modal="modal1031">31</button></div>
      </div>
    </div>
    </div>
 </div>
+<!--End of October Calendar-->
-<!--
+<!--Modal1001-->
-   <div class="fadein" class="main">
+   <div class="modal" id="modal1001">
-  <h3>Calendar</h3>
+    <div class="modal-content">
-  <p>From April 2019 to October 2019</p>
+      <div class="modal-header">October 1, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Adhesion functon test</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Adhesion test fluorescence checking using 4th floor plate reader machine. Result: Failed will elaborate later.</p>
+            <img class="img-fluid" style="display:block; margin-left: auto; margin-right: auto;"src="https://static.igem.org/mediawiki/2019/5/5c/T--UM_Macau--Lablog_1_Oct.jpg">
+            <br>
+            <p>Third functional test using 092819 1000ul activated bacteria. Put 5 ul of the yellow nanoparticle from Prof Xuanjun 30ppm into the 1000ul bacteria. Incubated at 25C for 1hr with shaking for 400 rpm. Started at 19:00.</p>
+            <br>
+            <p>Incubated new adhesion system bacteria for functional test. Picked all Colony #1. Prepared 5 mL overnight culture amp+, will subculture tomorrow to 20 mL culture (Need more bacteria concentration for the tests). Started the incubation at 22:30 at shaking incubator 37 C.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
    </div>
--->
+<!--End of Modal1001-->
-</div>
+<!--Modal1002-->
-<!--End of Calendar Content-->
+  <div class="modal" id="modal1002">
+    <div class="modal-content">
+      <div class="modal-header">October 2, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Incubation of bacteria culture</p>
+                <p>Induction of adhesion system</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a> <a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Subcultured 5 mL overnight cultures into 20 mL LB amp+ and incubated at 37 C for 3 hrs. Started incubation at 13:00. Saved the 5 mL overnight culture as uninduced samples. Will measure OD of both later, need OD=2 for activation and functional tests.</p>
+            <br>
+            <p>Induced Subcultured 20 mL adhesion system bacteria culture with 1 mM IPTG each (OPHT, OPHT2, OPHT3, OPHM, & OPHA). Incubated at 30 C for 4 hrs in Professor Xu's shaking incubator.</p>
+            <br>
+            <p>Update on the magnetization effect after incubation with ammonium ferric citrate of the BL-21 strain of E.coli. They can still be attracted to magnet until now since 092419. </p>
+            <br>
+            <p>Update on adhesion 20 mL culture, I will induce it overnight instead of 4 hrs. Will harvest them tomorrow morning instead.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1002-->
+<!--Modal1003-->
+  <div class="modal" id="modal1003">
+    <div class="modal-content">
+      <div class="modal-header">October 3, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Checked sequences of newly constructed plasmid </p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Checked the sequences of the newly constructed pet11a adhesion system plasmids. Conclusions are as follows：</p>
+<p>&nbsp;</p>
+<p>OPHT1 colony # 3 = Should be correct (Has a single point mutation but it's peak looks like a sequencing error)</p>
+<p>OPHT2 colony #1 = Correct and usable if needed</p>
+<p>OPHT3 = no correct or intact colony</p>
+<p>OPHA Colony #1 and #3 = Correct and usable if needed</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1003-->
+<!--Modal1004-->
+  <div class="modal" id="modal1004">
+    <div class="modal-content">
+      <div class="modal-header">October 4, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Sent sample for sequencing</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Send ftna samples that Julien extracted from 093019 to Tech dragon sequencing. </p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1004-->
+<!--Modal1005-->
+  <div class="modal" id="modal1005">
+    <div class="modal-content">
+      <div class="modal-header">October 5, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Functional test of the combination of tyrosinase and adhesion system</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Incubated 100ul bacteria with 50ul 1500U/ml tyrosinase and 1ul of 1%w/v yellow nanoparticle together for 4 hrs and 8hrs. Will check the fluorescence reading after these two time periods.</p>
+<p>&nbsp;</p>
+<p>Cultured 20 mL bacteria using glycerol stock of colony number 1 for OPHT, OPHT2,OPHT3, OPHM, OPHA1 with Amp+ and one for BL21 only without Amp.</p>
+<p>&nbsp;</p>
+<p>Current conclusions for adhesion system assay parameters: bacteria amount (Need OD=2), time of incubation with tyrosinase (4hrs, is the best I think), amount of tyrosinase (U/mL) (500U/ml or less), amount of nanoparticle (10ml/L or 10ppm is good for our experiment), centrifugation speed(3000xg 3 mins no nanoparticle pelleting), learning the plate reader machine(440 nm first filter and 480nm second filter), choosing the nanoparticle(1% w/v Yellow fluorescent particle from spherotech and red nanoparticle of Prof Xuan Jun), assay format in 96 well plate (As shown in the picture just now), Amount of bacteria and nanoparticle total volume (150 ul) so we can plate in 96 well.</p>
+<p>&nbsp;</p>
+<p>ligated pBAD24_NSP4 with fntA, and a negative control only has pBAD_NSP4 without insert then transformed them into DH5 alpha. Started incubation at 20:23.</p>
+<p>&nbsp;</p>
+<p>Cultured 18 tubes 20ml of bacteria from glycerol stock NSP4-Tyrosinase colony 3.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Current conclusion based on this fluorescence plate reader result is that after 4 hrs of incubation with tyrosinase there seems to be a significant decrease of supernatant fluorescence signal compared to our inactivated supernatant samples. Most notable OPHM.</p>
+              <img class="img-fluid" style="width: 100%; display: block; margin-left: auto; margin-right: auto; margin-top: 2%" src="https://static.igem.org/mediawiki/2019/0/00/T--UM_Macau--Lablog_5_Oct.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1005-->
+<!--Modal1006-->
+  <div class="modal" id="modal1006">
+    <div class="modal-content">
+      <div class="modal-header">October 6, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Induction of NSP4 tyrosinanse</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Induced with 5 concentrations of arabinose in 15 tubes of nsp4 tyrosinase bacteria overnight culture. Another 3 tubes are not induced with arabinose act as negative ctrl. The volume of arabinose added shown as below: </p>
+              <img class="img-fluid" style="width: 60%;margin-top: 2%; margin-bottom: 2%;" src="https://static.igem.org/mediawiki/2019/a/a4/T--UM_Macau--Lablog_6_Oct_1.jpg">
+            <br>
+            <p>Induced overnight OPHT1, OPHT2, OPHT3, OPHM and OPHA with 1 mM 40 ul of (500mM IPTG) per 20 mL culture and incubated in Professor Xu's shaking incubator 30 C for 4 hrs. Started ~approximately 12:08.</p>
+<p>&nbsp;</p>
+<p>Collected 4 hr incubation with 5 concentrations arabinose&rsquo;s bacteria culture.</p>
+<p>&nbsp;</p>
+<p>Nickel pull down for nsp4 4 hr arabinose induced (supernatant only), stored the pulled down protein in -80c protein samples box.</p>
+<p>&nbsp;</p>
+<p>Collected 8hr samples and centrifuging at 3000xg for 10 mins. Will separate supernatant into another tube.</p>
+<p>&nbsp;</p>
+<p>Incubated adhesion 100219 and 100519 bacteria with yellow nanoparticle started incubation at approx~ 21:00.</p>
+<p>&nbsp;</p>
+<p>Checked the construction of three extracted plasmids pbad24-fntA on 093019 by PCR and gel running.</p>
+<p>&nbsp;</p>
+<p>Nickel pull down for nsp4 8 hr arabinose induced (supernatant only), stored the pulled down protein in -80 protein samples box.</p>
+<p>&nbsp;</p>
+<p>Collected 12hr samples and centrifuging at 3000xg for 10 mins. Stored separated-supernatant into another tube and stored in -80c.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Nanoparticle Fluorescent signal of inactivated and activated supernatant of OPHT, OPHT2, OPHT3, OPHM, OPHA, Negative control, and Positive control.</p>
+              <img class="img-fluid" style="width: 100%; display: block; margin-left: auto; margin-right: auto; margin-top: 2%;" src="https://static.igem.org/mediawiki/2019/0/0c/T--UM_Macau--Lablog_6_Oct_2.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1006-->
+<!--Modal1007-->
+  <div class="modal" id="modal1007">
+    <div class="modal-content">
+      <div class="modal-header">October 7, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Incubation of bacteria containing FtnA plasmid</p>
+                <p>Functional test of adhesion system</p>
+                <p>Tyrosinase activity assay</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Incubated transformed bl-21 with FtnA plasmids. Diluted outgrowth solution 1:100 and plated 350 ul of the bacteria. Put it in Prof Xu&rsquo;s 37 degree incubator at 1:16 am.</p>
+<p>&nbsp;</p>
+<p>Incubated BL-21 with pBAD24_FtnA at 14:20 in Professor Xu's 37 C incubator. Only plated pBAD24_FtnA#1 and pBAD24_FtnA#2.</p>
+<p>&nbsp;</p>
+<p>Tyrosinase activity assay for 4,8,12 hours incubation with arabinose at 5 different concentrations. We cultured them at 30 degree accidentally.</p>
+<p>Tyrosinase activity assay reagents set-up:</p>
+<p>Sample Background Control (SBC) mix: add this into SBC wells</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>Assay buffer</p>
+</td>
+<td width="100%">
+<p>45ul x 18 = 810ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Enhancer</p>
+</td>
+<td width="100%">
+<p>5ul x 18 = 90ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Reaction mix: add this into assay background control, samples and positive control wells</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>Assay buffer</p>
+</td>
+<td width="100%">
+<p>35ul x 21 = 735ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Substrate</p>
+</td>
+<td width="100%">
+<p>10ul x 21 = 210ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Enhancer</p>
+</td>
+<td width="100%">
+<p>5ul x 21 = 105ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>Positive control well: 2ul positive control + 48ul assay buffer</p>
+<p>Assay background control: 50ul Assay buffer</p>
+<p>Plate:</p>
+<img class="img-fluid" style="width: 60%; margin-top: 2%; margin-bottom: 2%;" src="https://static.igem.org/mediawiki/2019/0/06/T--UM_Macau--Lablog_7_Oct.jpg">
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Results for the adhesion system functional test. Conclusion: The results are quite consistent.  Plate was read three times with the same machine and averaged all the reads as shown in the picture data set. Then I averaged each the three repeats for each samples and obtained final reads for each 100219 and 100519 samples and made the graph based on them. We can see a clear difference with the activated and inactivated one. Negative control (Wild type, just BL-21) is too close to our inactivated samples, not sure if the activated readings would be considered significant, but this negative control sample has been prepared along with the 100519 sample so it actually cannot act as a negative control for the 100219 samples but I just put it anyway. We need a third biological repeat for all these to have a final conclusion. Anyways, the data is very consistent, activated sample's readings are always lower than the inactivated ones for these first two biological repeats. </p>
+              <img class="img-fluid" style="width: 100%; display: block; margin-left: auto; margin-right: auto; margin-top: 2%;" src="https://static.igem.org/mediawiki/2019/5/56/T--UM_Macau--Lablog_7_Oct_2.jpg">
+              <img class="img-fluid" style="width: 80%; display: block; margin-left: auto; margin-right: auto; margin-top: 2%;" src="https://static.igem.org/mediawiki/2019/3/36/T--UM_Macau--Lablog_7_Oct_3.jpg">
+              <br>
+              <p>The results of plates after 1 hour measurement in the plate reader (30seconds interval, 37 degree incubation, OD 510nm) is saved.</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1007-->
+<!--Modal1009-->
+  <div class="modal" id="modal1009">
+    <div class="modal-content">
+      <div class="modal-header">October 9, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Nickel pull-down for NSP4 tyrosinase protein</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Incubated 15 tubes of 5ml nsp4 bacteria with 5 different arabinose concentration (2%, 0.2%, 0.02%, 0.002% and 0.0002%) at 12.30pm.</p>
+<p>&nbsp;</p>
+<table width="233">
+<tbody>
+<tr>
+<td width="116">
+<p>Concentrations of arabinose</p>
+</td>
+<td width="116">
+<p>Volume to add</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>2%</p>
+</td>
+<td width="116">
+<p>500ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>0.2%</p>
+</td>
+<td width="116">
+<p>50ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>0.02%</p>
+</td>
+<td width="116">
+<p>5ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>0.002%</p>
+</td>
+<td width="116">
+<p>0.5ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>0.0002%</p>
+</td>
+<td width="116">
+<p>0.05ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Collected nsp4 tyrosinase bacteria after 4hr incubation (at 4.30pm), centrifuged and keep the supernatant. Nickel pull down for 4hr incubation batch nsp4 bacteria&rsquo;s protein and stored in -80 fridge&rsquo;s bacteria box. (Because no space in protein sample box). Collected 8 hr incubation of different concentrations arabinose in nsp4 at 8.30pm. nickel pull down for 8hr incubation batch nsp4 bacteria&rsquo;s protein and stored in -80 fridge&rsquo;s bacteria box. (Cuz no space in protein sample box). Collected 12 hr arabinose incubation at 5 different concentrations of nsp4 bacteria at 12.30am . Centrifuged and kept the supernatant in -80 with a green holder.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1009-->
+<!--Modal1010-->
+  <div class="modal" id="modal1010">
+    <div class="modal-content">
+      <div class="modal-header">October 10, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Nickel pull down</p></td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Nickel pull down</p>
+            <br>
+            <p>Diluted protein samples by 10x and measure the protein concentration using BCA assay.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>Protein concentration:</p>
+              <img class="img-fluid" style="width: 100%; display: block; margin-left: auto; margin-right: auto; margin-top: 2%; margin-bottom: 2%;" src="https://static.igem.org/mediawiki/2019/6/63/T--UM_Macau--Lablog_10_Oct.jpg">
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1010-->
+<!--Modal1011-->
+  <div class="modal" id="modal1011">
+    <div class="modal-content">
+      <div class="modal-header">October 11, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien, Pinto</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td>
+              <p>Tyrosinase activity assay</p>
+              <p>Protein extraction</p>
+              <p>Western Blot</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a> <a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td>
+<p>Tyrosinase activity assay for 4,8,12 hours incubation with arabinose at 5 different concentrations. We cultured them at 37 degree this time.</p>
+<p>Tyrosinase activity assay reagents set-up:</p>
+<p>Sample Background Control (SBC) mix: We use only one SBC for each time point incubation of protein samples as we were running out of reagents.</p>
+<table width="275">
+<tbody>
+<tr>
+<td width="116">
+<p>Assay buffer</p>
+</td>
+<td width="159">
+<p>45ul x 3 = 135ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Enhancer</p>
+</td>
+<td width="159">
+<p>5ul x 3 = 15ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Reaction mix: add this into assay background control, samples and positive control wells</p>
+<table width="287">
+<tbody>
+<tr>
+<td width="116">
+<p>Assay buffer</p>
+</td>
+<td width="171">
+<p>35ul x 21 = 735ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Substrate</p>
+</td>
+<td width="171">
+<p>10ul x 21 = 210ul</p>
+</td>
+</tr>
+<tr>
+<td width="116">
+<p>Enhancer</p>
+</td>
+<td width="171">
+<p>5ul x 21 = 105ul</p>
+</td>
+</tr>
+</tbody>
+</table>
+<p>&nbsp;</p>
+<p>Positive control well: 2ul positive control + 48ul assay buffer</p>
+<p>Assay background control: 50ul Assay buffer</p>
+<p>&nbsp;</p>
+<p>Plates:</p>
+<img style="width: 50%; margin-top:2%; margin-bottom: 2%;" src="https://static.igem.org/mediawiki/2019/5/5c/T--UM_Macau--Lablog_11_Oct.jpg">
+<p>SDS-page gel loading sequence:</p>
+<p><u>First gel (OPHT-adhesion system):</u></p>
+<p>Ladder- Ladder- Control- OPHT1 - OPHT2 - OPHT3 - OPHM - OPHA1</p>
+<p>&nbsp;</p>
+<p><u>Second gel (ftNA):</u></p>
+<p>Ladder - Control - ftnA1.1 - ftnA1.2 - ftnA1.3 - ftnA2.1 - ftnA2.2 - ftnA2.3</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>In the functional test results page</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1011-->
+<!--Modal1012-->
+  <div class="modal" id="modal1012">
+    <div class="modal-content">
+      <div class="modal-header">October 12, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>ftnA functional test</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Centrifuged the second bench ftnA test samples and resuspend in LB broth. Measure the OD value. Begin magnet attraction test at 9:30 and will check their states every hour.</p>
+              <div class="row" style="width: 60%; margin-top: 2%; margin-left: auto; margin-right: auto; display: block;">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/d/d4/T--UM_Macau--Lablog_12_Oct_1.jpg">
+              </div>
+              <div class="row" style="width: 60%; margin-top: 2%; margin-left: auto; margin-right: auto; display: block;">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/1/12/T--UM_Macau--Lablog_12_Oct_2.jpg">
+              </div>
+              <div class="row" style="width: 60%; margin-top: 2%; margin-left: auto; margin-right: auto; display: block;">
+                <img class="img-fluid" src="https://static.igem.org/mediawiki/2019/e/ec/T--UM_Macau--Lablog_12_Oct_3.jpg">
+              </div>
+            <br>
+            <p>Transformation of pBAD24-FtnA #2 into seven strains which are Stbl3, DH5 alpha, BL-21 DE3, BL-21 Star, BL-21 Rosetta, BL-21 pLys, TOP10. Started incubation at 13:00.</p>
+            <br>
+            <p>Reset pBAD24-FtnA 1.1, 1.2, 2.1, 2.2 attraction test with only one magnet under it at 14:50.</p>
+            <br>
+            <p>Induced 5 tubes 5ml bacteria pBAD24-FtnA 1.1 with 1ml 100mM ferric ammonium citrate at 16:20.</p>
+            <br>
+            <p>101219: Incubated 20 mL BL-21 NSP4 tyrosinase on 37 degree for adhesion system functional test with our own enzyme. </p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1012-->
+<!--Modal1013-->
+  <div class="modal" id="modal1013">
+    <div class="modal-content">
+      <div class="modal-header">October 13, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Adhesion system functional test</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Started adhesion system functional test. Incubated adhesion system bacteria with yellow nanoparticle. Would collect the samples at 4 different time points, 1 hr, 2 hr, 3 hr and 4 hr and compare the difference in the relative fluorescence.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1013-->
+<!--Modal1014-->
+  <div class="modal" id="modal1014">
+    <div class="modal-content">
+      <div class="modal-header">October 14, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Bacteria culture, Incubate with antibody for immunoblotting membrane</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf">Plasmid Construction</a> <a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Incubated 5ml culture of  FtnA-Stbl3,Dh5alpha,BL-21(De3),BL-21(star), BL-21(plys), TOP10, BL-21(Rosetta), And Bl21(WT) at 37 degree shaking incubator started 14:39.</p>
+              <br>
+              <p>Pinto made SDS gel and store in Buffer. Measured protein concentration of FtnA protein samples and stored in -80C.</p>
+              <br>
+              <p>Autoclaved some tips blue and yellow tips and 1.5 mL centrifuge tubes.</p>
+              <br>
+              <p>Incubated FtnA samples and RpoB internal control with anti-flag and anti-rabbit primary antibody. Put 5 mL and 3 mL respectively of the antibodies. Put in 4 degree in Henry’s fridge at 20:20. </p>
+              <br>
+              <p>Incubated 6 20mL culture of OPHT, OPHT2,OPHT3, OPHM, OPHA and WT bl-21 all picked from glycerol stock colony #1 in 37 degree shaking incubator overnight(start 21:00) Has white tape mark, will induce it with 1mM IPTG for 4 hrs after 12 hrs (9:00). For time dependent functional test with yellow fluorescent nanoparticle and Professor Xuan Jun’s nanoparticle. </p>
+              <br>
+              <p>Incubated 5x 20 mL culture of NSP4_BL21 subcultured from yesterday’s overnight culture(1:100 ratio) at 37C started at (21:00) . Will induce later with 0.2% arabinose after 3hrs (00:00). And let it produce tyrosinase into the supernatant for overnight(12-16 hrs) Has green tape mark. This is for testing our own secreted tyrosinase whether it would be able to activate our capture system bacteria.</p>
+              <br>
+              <p>Incubated 6x 5mL tubes of OPHT,OPHT2,OPHT3,OPHM, OPHA and negative ctrl at 37 C has yellow tape mark started (21:00). Picked from glycerol stock colony #1. Will incubate overnight and induce tomorrow after 16 hrs with IPTG and incubate in 30C for 4 hrs (12:00). </p>
+              <br>
+              <p>Incubated 7x 5mL tubes for each bacteria strains we had cultured overnight FtnA-Stbl3,Dh5alpha,BL-21(De3),BL-21(star), BL-21(plys), TOP10, BL-21(Rosetta), And Bl21(WT) at 37 degree shaking incubator started 21:00. Has red tape. Subcultured 1:100 ratio will induce with 0.2% arabinose later after 3 hrs (00:00) and incubate for 12-16 hrs. These samples are for protein concentration check for FtnA based on different strains. Will extract proteins tomorrow and measure protein concentration along with the preparation of protein samples for western blot running on Wednesday.</p>
+              <br>
+              <p>Induced earlier log picked strains incubated this morning from 2:39 5ml cultures of  FtnA-Stbl3,Dh5alpha,BL-21(De3),BL-21(star), BL-21(plys), TOP10, BL-21(Rosetta), And Bl21(WT) with 0.2% arabinose (50 ul of 20% arabinose) and stored in 37 degree shaking incubator has purple tape. Started at 21:42. Will incubate overnight for 12 hrs. Then would put with ~20 mM ammonium ferric Citrate 1 mL each and incubate for 24 hrs for magnetization test on Wednesday morning 9:00 am.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1014-->
+<!--Modal1015-->
+  <div class="modal" id="modal1015">
+    <div class="modal-content">
+      <div class="modal-header">October 15, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Western blot</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Induced red and green taped samples from previous logs with 0.2% arabinose at 00:44 and put in the 37 C shaking incubator. Plan to incubate for 12-16 hrs (Probably 16 hrs). </p>
+              <br>
+              <p>Induced yellow tape marked tubes and white marked tape tubes with 1mM IPTG at 12:00, put them in 30C shaking incubator of Professor Xu's lab.</p>
+              <br>
+              <p>Washed the membrane with tbst 3 times and incubated with anti-rabbit 2nd antibody at 12.40pm</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1015-->
+<!--Modal1016-->
+  <div class="modal" id="modal1016">
+    <div class="modal-content">
+      <div class="modal-header">October 16, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Julien</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Western blot</p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Incubated purple taped samples from 101419 with ~20 mM ferric ammonium citrate. At 37 C starting 00:49.</p>
+              <p>Run SDS-Page gel. Loading order as below:</p>
+              <p><u>First gel</u></p>
+              <p>Ladder-Ladder-Control-2%-0.2%-0.02%-0.002%-0.0002%</p>
+              <p>&nbsp;</p>
+              <p><u>Second gel</u></p>
+              <p>Ladder-BL-21WT-Rosetta-BL-21(star)-TOP10-BL-21(plys)-Dh5alpha-BL-21(De3)</p>
+              <p>&nbsp;</p>
+              <p>Incubated membranes with primary antibody for overnight. (Anti-flag for 19kda ftna, Anti-rpob for 150kda internal ctrl).</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1016-->
+<!--Modal1017-->
+  <div class="modal" id="modal1017">
+    <div class="modal-content">
+      <div class="modal-header">October 17, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Immunofluorescence</p>
+                <p></p>
+                <p></p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf">Protein Expression</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Incubated immunofluorescence slide samples with histidine primary antibody for 12-16 hrs. Put in 4 degree in Henry’s fridge. </p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modals-->
+<!--Modal Template-->
+  <div class="modal" id="modal1018">
+    <div class="modal-content">
+      <div class="modal-header">October 18, 2019
+        <span class="modal-close closeicon">&times;</span></div>
+      <div class="modal-body">
+        <table>
+        <!--Performers-->
+          <tr>
+            <th>Performers</th>
+            <td><p>Philip, Renee</p></td>
+          </tr>
+        <!--Experiments-->
+          <tr>
+            <th>Experiments</th>
+            <td><p>Cell viability testing</p>
+                <p></p>
+                <p></p>
+              </td>
+          </tr>
+        <!--Protocols-->
+          <tr>
+            <th>Protocols</th>
+            <td><a target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf">Functional Test</a></td>
+          </tr>
+        <!--Records-->
+          <tr>
+            <th>Records</th>
+            <td><p>Started incubation of bacteria cell viability with ZnO and AgNP at 04:30. Will test the following time points (Hrs) or OD600 (2, 4,6,8,10,12,14,16). 6:30, 8:30, 10:30, 12:30, 14:30, 16:30, 18:30 are the time points. I would need help getting the absorbance values for the 10:30, 12:30, 14:30 timepoints. </p>
+            <br>
+            <p>Incubated 20 mL overnight culture OPHt,OpHT2, OPHT3, OPHM, OPHA, -ctrl in 37 C shaking incubator started 17:00.</p>
+            </td>
+          </tr>
+        <!--Results-->
+          <tr>
+            <th>Results</th>
+            <td>
+              <p>None</p>
+            </td>
+          </tr>
+        </table>
+      </div>
+      <div class="modal-footer"><button class="link modal-close">Close</button></div>
+    </div>
+  </div>
+<!--End of Modal1017-->
+</div>
+<!--End of Calendar Content-->
 <!--Protocol Content-->
 <div id="Protocols" class="tabcontent">
-   <h3>Protocols</h3>
+   <div class="row" style="padding:10%; text-align: center">
-  <p>Protocols used in the Project SANCE</p>
+    <div class="col-sm-4">
+      <a  target=_blank href="https://static.igem.org/mediawiki/2019/c/c0/T--UM_Macau--Plasmid_Construction.pdf"><img class="img-fluid" src="https://static.igem.org/mediawiki/2019/6/63/T--UM_Macau--documenticon.png"></a>
+      <h3>Plasmid Construction</h3>
+    </div>
+    <div class="col-sm-4">
+      <a  target=_blank href="https://static.igem.org/mediawiki/2019/a/ac/T--UM_Macau--Protein_Expression.pdf"><img class="img-fluid" src="https://static.igem.org/mediawiki/2019/6/63/T--UM_Macau--documenticon.png"></a>
+      <h3>Protein Expression</h3>
+    </div>
+    <div class="col-sm-4">
+      <a  target=_blank href="https://static.igem.org/mediawiki/2019/b/b9/T--UM_Macau--Functional_Test.pdf"><img class="img-fluid" src="https://static.igem.org/mediawiki/2019/6/63/T--UM_Macau--documenticon.png"></a>
+      <h3>Functional Test</h3>
+    </div>
+  </div>
 </div>
 <!--End of Protocol Content-->
-<!--Lablogs Content-->
-<div id="Lablogs" class="tabcontent">
-  <h3>Lab Logs</h3>
-  <p>A full lablogs with all details and used protocols</p>
-</div>
-<!--End of Lablogs Content-->
 </div>

Difference between revisions of "Team:UM Macau/Notebook"

Latest revision as of 04:16, 17 December 2019

Notebook

May

June

July

August

September

October

Plasmid Construction

Protein Expression

Functional Test