Revision as of 16:30, 20 October 2019

<!DOCTYPE html> Cleaning factory: methotrexate · Benchling

Cleaning factory: methotrexate

Project: Cleaning factory

Authors: iGEM_GO_Paris_saclay

Thursday, 11/7/2019

Aim: We successively induce the expression of the MTX degradation pathway and then of nuclease_gp3 . Thereafter bacteria were incubated in the presence of MTX. We monitored MTX biotransformation by HPLC analysis. MTX should be degrated as if there was no nuclease induction.

For our cleaning factory, our 2 plasmids must not have the same replication origin to not lose one of them. Then, we amplify the replication origin 15A in pZA-15A31 and pZA-15A34 in order to put it in pBAD24 instead of the ColE1. For this, we use the Gibson assembly method.

●

Correction of the replication origin of pZA-15A31 and pZA-15A34 : Marie_Natacha

Nanodrop : (from Philippe's tubes)

pZA-15A31 -> 0,161 ng/µL ; A260 0,0032 ; 260/280 -0,32 ; 260/280 -0,32.

pZA-15A34 -> negative concentration.....

○

PCR protocol :

2,5 µL Ori_15A_F

2,5 µL Ori_15A_R

2 µL pZA-15A31 or pZA-15A34

25 µL Master Mix 2X

18 µL H2O mQ

-> total 50 µL

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-7- Ori_15A_F	GACCTAGGGGATATATTCCGCT	?
3 3	2019GO-8- Ori_15A_R	GATATCGAGCTCGCTTGGAC	?

Primer sequences for changing the replication origin of pZA31 or 34

○

PCR program:

98°C/30sec

25 cycles : 98°C/10sec

65°C/30sec

72°C/30sec

72°C/2min

infinite 15°C

○

agarose 1% gel, 100V 25min. Results :

image.png

○

Heatshock plasmid transformation (Natacha)

Let DH5alpha on ice for 30 min

Add plasmid PZA34 and PZA31M2N (a few µL) in your bacterial solution and let it on ice for 30 min

Heatshock at 42°C for 1 min

Put the solution back in ice for 2 min

Add 1 mL LB (+ Glu) and put the solution at 37°C for 1 h with shaking

Spread out transformants on agar plates (50 µL, 100 µL and 100 µL concentrate)

Put the plates at 37°C overnight

Friday, 12/7/2019

●

Plasmid extraction of pZA31, pZA34 and pSB1C3 : Arnaud

Use of Macherey-Nagel kit "DNA and RNA purification" protocol page 18 "Low copy"

3 extractions per plasmids pZA31 and pZA34 and 2 extractions for pSB1C3

Analysis with Nanodrop :

	A A	B B	C C	D D
1 1	Plasmid	ng/uL	260/230	260/280
2 2	pSB1C3 (1)	52.4	2.39	2.04
3 3	pSB1C3 (2)	66.84	2.21	2.01
4 4	pZA34 (1)	10.57	1.09	2.56
5 5	pZA34 (2)	13.08	1.15	2.21
6 6	pZA34 (3)	12.34	1.61	2.51
7 7	pZA31 (1)	12.74	2.10	2.70
8 8	pZA31 (2)	14.35	1.40	2.63
9 9	pZA31 (3)	13.79	2.08	2.61

Table4

●

PCR for Gibson assembly of pBAD24A1/pBAD24_Moclo: Kim

PCR of plasmid pBAD24 with or without the A1 gene.

Expected size : pBAD24_A1 = 5,5 kB

pBAD24_Moclo = 3,9 kB

A1 = 1,6 kB

Primers: pBAD24_F (9) and pBAD24_R (10)

Tm = 62°C

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-9- ori15A_pBAD24_v2_F	gtccaagcgagctcgatatcgagcctatggaaaaacgcca
3 3	2019GO-10- ori15A_pBAD24 _v2_R	agcggaatatatcccctaggtcgggattttggtcatgagattatc

Primer sequences for Gibson assembly to remove the replicative origins

Follow the protocol "PCR Q5 HF"

2 reactions per plasmid (50µL)

PCR program:

(30 cycles)

98°C 30 sec

98°C 10 sec

62°C 3 min

72°C 3 min

72°C 2 min

5 µL of each reaction were migrated on agarose 1%. No fragments were detected. The PCR needs to be done again.

Monday, 15/7/2019

●

PCR of pZA31, pZA34, pBAD24_A1 and pBAD24_Moclo : Arnaud_Abdel

For pZA31 and pZA34, Primers : Ori 15 A_F and Ori 15 A_R

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-7- Ori_15A_F	GACCTAGGGGATATATTCCGCT	?
3 3	2019GO-8- Ori_15A_R	GATATCGAGCTCGCTTGGAC	?

Table5

For pBAD24_A1 and pBAD24_Moclo, Primers : pBAD24_F and pBAD24_R

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-9- ori15A_pBAD24_v2_F	gtccaagcgagctcgatatcgagcctatggaaaaacgcca
3 3	2019GO-10- ori15A_pBAD24 _v2_R	agcggaatatatcccctaggtcgggattttggtcatgagattatc

Table6

For 1 PCR , Vf = 90 µL

■

2.5 µL per primers 10mM

■

1 µL template

■

25 µL Master Mix (Enzyme, dNTPs, Buffer)

■

18 µL Water

The 4 PCR are done simultaneously with the following program:

I.

98°C - 1 min

II.

98°C - 10 s

III.

62°C - 30 s

IV.

72°C - 165 s

V.

Repeat from step 2 (30 times)

VI.

72°C - 2 min

VII.

8°C - Infinity

The products are analysed by electrophoresis

Gel PCR 15.07.jpg

Wednesday, 17/7/2019

Gibson cloning, substitute replication origin ColE1 with A15 : Hugues

○

Extraction on gel of pBAD24-A1 (without Ori ColE1) with the kit DNA extraction Macherey-Nagel

○

Digestion of template plasmids with DpnI, 0,5µL of enzyme (37°C 1h then 80°C 20min)

○

PCR clean up Macherey-Nagel (elution in 30µL NE buffer)

○

Nanodrop pBAD24-A1(without Ori ColE1) = 1,5 ng/µL

Thursday, 18/7/2019

●

Nanodrop and Gibson assembly of A15 (1) and A15(4) (Kim and Hugues):

1µl DpnI 1h at 37°C. Then 20 minutes at 80°C. PCR clean-up.

Nanodrop concentration:

A15 (1) = 100 ng/µl

A15 (4) = 100 ng/µl

Gibson:

1h with Gibson mix at 50°C. 1:1 ratio --> 4µl of plasmid + 1 µl of (1) and (4) each diluted at 1/10.

●

PCR on plasmid pBAD24-MoClo in order to remove the replication origin: Hugues

PCR protocol:

25µL buffer Q5 2X

2,5µL primer 9

2,5µL primer 10

1µL pBAD24 MoClo

10µL solution Q

9µL H2O

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-9- ori15A_pBAD24_v2_F	gtccaagcgagctcgatatcgagcctatggaaaaacgcca
3 3	2019GO-10- ori15A_pBAD24 _v2_R	agcggaatatatcccctaggtcgggattttggtcatgagattatc

Primer sequences for gibson assembly in order to remove the origin of pBAD24-Moclo

PCR program used:

98°C 30sec

98°C 10sec

62°C 20sec *5

72°C 2min

98°C 10sec

72°C 20sec *25

72°C 2min

Monday, 22/7/2019

●

PCR colony on bacteria transformed with Gibson products : Arnaud

PCR program:

I.

98°C 5min

II.

98°C 30sec

III.

54°C 30sec

IV.

72°C 1min

V.

Repeat from step 2 (30times)

VI.

72°C 10min

VII.

8°C Infinity

PCR protocol :

Vf = 25µL

dXTPs 5 mM 1µL

Oligos (7 et 8) 1µL/oligo

Matrice 1µL

Enzyme DreamTaq 0.5µL

Tampon DreamTaq 10X 2.5µL

H2O 18µL

Results : Analysis on eletrophoresis showed no success with the PCR

●

Gibson : Arnaud

Tomorrow:

DpnI digestion

PCR clean up

Transformation into competent cells

Tuesday, 23/7/2019

PCR for Gibson assembly of pBAD24A1/pBAD24A1 without Ori : Colombe

PCR of plasmid pBAD24A1 with or without the ori.

Expected size : pBAD24_A1 = 5,5 kB = 5575 B

Primers: pBAD24_F (9) and pBAD24_R (10)

Tm = 62°C

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-9- ori15A_pBAD24_v2_F	gtccaagcgagctcgatatcgagcctatggaaaaacgcca
3 3	2019GO-10- ori15A_pBAD24 _v2_R	agcggaatatatcccctaggtcgggattttggtcatgagattatc

Primer sequences for gibson assembly in order to remove the origin of pBAD24-Moclo

Follow the protocol "PCR Q5 HF"

2 reactions per plasmid (50µL) : 4 reactions in total

PCR program:

(30 cycles)

98°C 30 sec

98°C 10 sec

62°C 3 min _ 5 cycles

72°C 3 min _ 25 cycles

72°C 5 min

RESULTS :

wells 1 & 2 : pBAD24_A1 with ori => no results because of empty eppendorf ?

wells 3 & 4 : pBAD24_A1 without ori from gel extraction of the previous PCR (Arnaud ?)

IMG_8718.JPG

(5 µL of each reaction were migrated on agarose 1%.)

Then DNA extraction from agarose gel with gel extraction's kit of NucleoSpin^R Gel and PCR clean-up, macherey-nagel

Seen NucleoSpin Macherey-Nagel protocol

Wednesday, 31/7/2019

●

PCR pBAD-A1 (without ori colE1) : Laetitia et Mahnaz

Oligos 9 et 10

	A A	B B	C C
1 1	Primer name	Primer sequence	Tm °C
2 2	2019GO-9- ori15A_pBAD24_v2_F	gtccaagcgagctcgatatcgagcctatggaaaaacgcca
3 3	2019GO-10- ori15A_pBAD24 _v2_R	agcggaatatatcccctaggtcgggattttggtcatgagattatc

Primer sequences for gibson assembly in order to remove the origin of pBAD24-Moclo (2)

DNA : pBAD24-A1 118ng (nd, PCR1) or 11,8ng (diluted 10x, PCR2) or 1,18ng (diluted 100x, PCR3)

Enz Q5

PCR program :

98°C/30sec

5 cycles : 98°C/10sec

52°C/30sec

72°C/3min

20 cycles : 98°C/10sec

72°C/30sec

72°C/3min

72°C/2min

Results :

Friday, 2/8/2019

●

Gibson Assembly of pBAD-A1 (without origin) with origin 15A : Laetitia

3 assemblies are done. The third one is a negative controle with only the pBAD-A1 without origin PCR fragment

Assembly 1 :

1.

pBAD-A1 without origin PCR fragment 2,5 µL of PCR3 (about 300ng) (for PCR details, see July 31st)

2.

PCR Product origin 15A diluted 7 times 2,5µL

3.

Gibson assembly mix 15 µL

Final volume = 20 µL

Assembly 2 :

1.

pBAD-A1 without origin PCR fragment 2,5 µL of PCR1 (about 300ng) (for PCR details, see July 31st)

2.

PCR Product origin 15A diluted 7 times 2,5µL

3.

Gibson assembly mix 15 µL

Final volume = 20 µL

Assembly 3 (control) :

1.

pBAD-A1 without origin PCR fragment 2,5 µL of PCR1 (about 300ng) (for PCR details, see July 31st)

2.

H2O 2,5µL

3.

Gibson assembly mix 15 µL

Final volume = 20 µL

Then it is put at 50°C for one hour (PCR machine) and store in ice for transformation later on.

●

Heatshock plasmid transformation (with Golden gate and Gibson done above) : Laetitia

1.

50 µL of ultra-competent cells DH5alpha E. coli +

2 µL from Gibson assembly of pBAD-A1 (without origin) with origin 15A n°1 or

2 µL from Gibson assembly of pBAD-A1 (without origin) with origin 15A n°2 or

2 µL from Gibson assembly of pBAD-A1 (without origin) without origin 15A (control) n°3 or

2 µL from Golden Gate of pBADMoclo with A1 and RBS_Rdrp or

2 µL from Golden Gate of pBADMoclo alone (control)

2. 30min on ice

3. Heat shock 90s at 42°C

4. 15min on ice and then add 1mL of LB and put tubes at 37°C for 1h

5. Spread cells on LB+Amp+Glu and cells are incubated at 37°c for 24h

Results :

●

With Gibson products : No colony to be seen on any plate. Transformation didn't work. => Fail

●

With Golden Gate : No colony for the transformants control (pBAD24_A1). Colonies available for the transformants pBAD24_A1_RBS_Rdrp on the plates. => Success

Thursday, 8/8/2019

●

PCR of pBAD24-A1, pBAD24-T7 and pBAD24-MoClo for Gibson assembly (Clara)

Gibson assembly was designed in order to make all our plasmid origins compatible between each of them (pBAD24 and pZA31-Cpg2)

For this PCR the master mix of Q5 was used, for more details, see the protocol "PCR Q5 HF". PCR products have been amplified everywhere except for the Ori part.

The PCR program used was the following:

	A A	B B	C C
1 1	STEP	TEMP	TIME
2 2	Initial denaturation	98°C	30 sec
3 3	35 cycles	98°C	30 sec
4 4		60°C	30 sec
5 5		72°C	3 min
6 6	Final extension	72°C	10 min
7 7	Hold	4-10°C

Table1

However, we observed a smear after electrophoresis, so the PCR was done again the next day with diluted DNA.

The photo of this gel wasn't saved.

Friday, 9/8/2019

●

PCR of pBAD24-A1, pBAD24-T7 and pBAD24-MoClo for Gibson assembly (Clara)

Gibson assembly was designed in order to make all our plasmid origins compatible between each of them (pBAD24 and pZA31-Cpg2)

For this PCR the master mix of Q5 was used, for more details, see the protocol "PCR Q5 HF". PCR products have been amplified everywhere except for the Ori part.

The PCR program used was the following:

	A A	B B	C C
1 1	STEP	TEMP	TIME
2 2	Initial denaturation	98°C	30 sec
3 3	5 cycles	98°C	30 sec
4 4		60°C	30 sec
5 5		72°C	3 min
6 6	Final extension	72°C	10 min
7 7	20 cycles	98°C	10 sec
8 8		72°C	30 sec
9 9		72°C	3 min
10 10	Hold	4-10°C
11 11

Table2

DNAs were diluted 10 and 100 times pBAD-A1:? and pBAD-MoClo: 118ng/µl) except for pBAD24-T7 (plasmid concentration was 16ng/µl).

Results:

PCR pbadmoclo et pbad A1 T7.pdf

pBAD24-MoClo wasn't amplified, maybe a pipetting error. Have to do the PCR for this plasmid again, the expected size should have been 3946 pb.

Monday, 12/8/2019

●

PCR of pBAD24-MoClo for Gibson assembly (Clara)

Gibson assembly was designed in order to make all our plasmid origins compatible between each of them (pBAD24 and pZA31-Cpg2)

For this PCR the master mix of Q5 was used, for more details, see the protocol "PCR Q5 HF". PCR products have been amplified everywhere except for the Ori part.

The PCR program used was the following:

	A A	B B	C C
1 1	STEP	TEMP	TIME
2 2	Initial denaturation	98°C	30 sec
3 3	5 cycles	98°C	30 sec
4 4		60°C	30 sec
5 5		72°C	3 min
6 6	Final extension	72°C	10 min
7 7	20 cycles	98°C	10 sec
8 8		72°C	30 sec
9 9		72°C	3 min
10 10	Hold	4-10°C
11 11

Table3

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      </style><div><div><div class="printable-container hide-print-links" data-printable-left-margin="38"><div class="printable-container-boundingBox"><div><div class="printable-page-container"><div class="noteEditor mediocre" tabindex="-1"><div></div><div class="noteEditor-container"><div class="u-cursor-text noteEditor-catcher u-disable-margin-collapse dnd-drop-container"><div class="noteEditor-contentWrapper" data-mediocre-item-whitespace="1"><h1 class="mediocre-titleEditor"><span class="mediocre-titleEditor-header"><span class="mediocre-titleEditor-header-id u-faded"> </span></span><div class="mediocre-titleEditor-titleEditable editable selectable is-locked" contenteditable="false" data-allow-multiline="false" data-cid="notetitleeditor" data-index="0">Cleaning factory: methotrexate</div></h1><div><div><div><b>Project:</b> Cleaning factory</div><div><b>Authors:</b> iGEM_GO_Paris_saclay</div></div></div><div class="editor" data-editor="notes"><div class="hiddenFocusEditable js-focusBearer" contenteditable="true" tabindex="-1"></div><div class="daySeparator" data-scroll-id="entryDay842"><div class="daySeparator-inputWrapper"><input class="daySeparator-inputWrapper-hiddenInput" type="input"/></div><span class="daySeparator-date">Thursday, 11/7/2019</span><div class="daySeparator-line"></div></div><div class="mediocre-item is-text" data-scroll-id="note-part886"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part886" data-index="0" style="margin-left: 35px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part887"><div class="mediocre-noteEditable multi-editable is-header2"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part887" data-index="1" style="margin-left: 35px;"><u><span style="color:#000000">Aim</span></u><span style="color:#000000">: W<span style="background-color:#FFFFFF">e successively induce the expression of the MTX degradation pathway  and then of </span><i><span style="background-color:#FFFFFF">nuclease_gp3</span></i><span style="background-color:#FFFFFF"> . Thereafter bacteria were incubated in the presence of MTX. We monitored MTX biotransformation by HPLC analysis.</span> MTX should be degrated as if there was no nuclease induction.</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part888"><div class="mediocre-noteEditable multi-editable is-header2"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part888" data-index="2" style="margin-left: 35px;"><span style="color:#000000">For our cleaning factory, our 2 plasmids must not have the same replication origin to not lose one of them. Then, we amplify the replication origin 15A in pZA-15A31 and pZA-15A34 in order to put it in pBAD24 instead of the ColE1. For this, we use the Gibson assembly method.</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part889"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part889" data-index="3" style="margin-left: 35px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part890"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"><span>●</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part890" data-index="4" style="margin-left: 35px;"><b>Correction of the replication origin of pZA-15A31 and pZA-15A34 : Marie_Natacha</b></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part891"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part891" data-index="5" style="margin-left: 35px;">Nanodrop : (from Philippe's tubes) </div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part892"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part892" data-index="6" style="margin-left: 35px;">pZA-15A31 -&gt; 0,161 ng/<span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span>L ; A260 0,0032 ; 260/280 -0,32 ; 260/280 -0,32.</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part893"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part893" data-index="7" style="margin-left: 35px;">pZA-15A34 -&gt; negative concentration.....</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part894"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part894" data-index="8" style="margin-left: 35px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part895"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"><span>○</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part895" data-index="9" style="margin-left: 70px;">PCR protocol : </div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part896"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part896" data-index="10" style="margin-left: 70px;">2,5 <span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span><span style="color:#344043"><span style="background-color:#FFFFFF">L </span></span><span style="color:#344043"><span style="background-color:#FFFFFF">Ori_15A_F</span></span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part897"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part897" data-index="11" style="margin-left: 70px;"><span style="color:#344043"><span style="background-color:#FFFFFF">2,5 </span></span><span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span><span style="color:#344043"><span style="background-color:#FFFFFF">L </span></span><span style="color:#344043"><span style="background-color:#FFFFFF">Ori_15A_R</span></span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part898"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part898" data-index="12" style="margin-left: 70px;">2 <span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span><span style="color:#344043"><span style="background-color:#FFFFFF">L </span></span><span style="color:#344043"><span style="background-color:#FFFFFF">pZA-15A31</span></span><span style="color:#344043"><span style="background-color:#FFFFFF"> or </span></span><span style="color:#344043"><span style="background-color:#FFFFFF">pZA-15A34</span></span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part899"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part899" data-index="13" style="margin-left: 70px;">25 <span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span><span style="color:#344043"><span style="background-color:#FFFFFF">L Master Mix 2X</span></span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part900"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part900" data-index="14" style="margin-left: 70px;">18 <span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span><span style="color:#344043"><span style="background-color:#FFFFFF">L H2O mQ</span></span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part901"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part901" data-index="15" style="margin-left: 70px;">-&gt; total 50 <span style="color:#344043"><span style="background-color:#FFFFFF">µ</span></span><span style="color:#344043"><span style="background-color:#FFFFFF">L </span></span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part902"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part902" data-index="16" style="margin-left: 70px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part903"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part903" data-index="17" style="margin-left: 70px;"></div></div></div><div class="mediocre-item is-captionedTable" data-scale-for-print="print1906" data-scroll-id="note-part904"><div class=""><div><div class="hiddenFocusEditable editable" contenteditable="true" data-cid="note-part904-item" data-formattable="true" data-index="18" data-subindex="item"></div><div class="mediocre-captionEditable multi-editable mediocre-captionEditable--table"><div class="mediocre-captionEditable-indent"><div class="mediocre-captionEditable-grid"><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell"></div><div class="mediocre-captionEditable-cell mediocre-captionEditable-component" style="width: 100%;"><div class="note-item-wrapper"><div class="mediocre-tableEditable printable-container-scaleForPrint" data-formattable="true" data-index="18" data-printable-layout-width="763" data-subindex="cell"><div></div><div class="hiddenFocusEditable js-focusBearer" contenteditable="true" tabindex="-1"></div><div class="hiddenFocusEditable editable hiddenFocusEditable--fullTable" contenteditable="true" data-cid="note-part904-table" data-formattable="true" data-index="18" data-subindex="table"></div><div class="u-position-relative mediocre-tableEditable-scrollAreaWrapper" style="margin-left: 0px; margin-right: 0px;"><div class="mediocre-tableEditable-scrollArea" style="padding-top: 47px;"><div class="mediocre-tableEditable-tableWrapper" style="padding-left: 70px;"><div class="mediocre-tableEditable-border multi-editable" style="left: 64px; top: -43px; bottom: 4px; width: 755px; min-width: 12px;"></div><div class="u-position-relative u-flex-display"><table class="mediocre-tableEditable-table mediocre-tableEditable-table--main" style="width: 743px;"><colgroup><col style="width: 45px;"></col><col data-id="0" style="width: 207px;"></col><col data-id="1" style="width: 365px;"></col><col data-id="2" style="width: 126px;"></col></colgroup><tbody><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--header"></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">A</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">A</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">B</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">B</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">C</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">C</span><div class="u-flex-display"></div></div></div></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">1</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">1</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-0-0" data-namespace="table-ns1831"><span>Primer name</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-0-1" data-namespace="table-ns1831"><span>Primer sequence</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-0-2" data-namespace="table-ns1831"><span>Tm °C</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">2</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">2</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-1-0" data-namespace="table-ns1831"><span>2019GO-7- Ori_15A_F </span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-1-1" data-namespace="table-ns1831"><span>GACCTAGGGGATATATTCCGCT
 </span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-1-2" data-namespace="table-ns1831"><span>?</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">3</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">3</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-2-0" data-namespace="table-ns1831"><span>2019GO-8- Ori_15A_R         </span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-2-1" data-namespace="table-ns1831"><span>GATATCGAGCTCGCTTGGAC</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part904-2-2" data-namespace="table-ns1831"><span>?</span></td></tr></tbody><tfoot></tfoot></table><div class="mediocre-tableEditable-fillerTableWrapper" style="margin-right: 10px;"><table class="mediocre-tableEditable-fillerTable" style="display: none;"><tbody><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr></tbody></table></div></div></div></div></div><div class="mediocre-tableEditable-titleBarWrapper"><div class="mediocre-tableEditable-titleBar" style="height: 37px; padding-left: 70px;"><div class="mediocre-tableEditable-tableName u-pointer"><div class="u-flex-grow u-overflow-hidden u-verticalCenterItems"><div class="u-ellipsis-text"><span class="u-ellipsis-text">Primer sequences for changing the replication origin of pZA31 or 34</span></div></div><div class="u-flex-none"></div></div><div class="u-flex-none u-tiny-spacing-left u-verticalCenterItems table-floater"><svg class="svgIcon workflowTable-tableCollapser u-pointer"><use xlink:href="#chevron-up_c0f0f77c91b0d9072b0d8255f9f54e3c"></use></svg></div></div></div></div></div></div><div class="mediocre-captionEditable-cell"></div></div><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell"></div><div class="mediocre-captionEditable-cell"><div class="mediocre-captionEditable-caption--bottom mediocre-captionEditable-caption editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="true" data-cid="note-part904" data-index="18" style="max-width: 763px; margin-left: 70px;">
-</div></div><div class="mediocre-captionEditable-cell"></div></div></div></div></div></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part905"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part905" data-index="19" style="margin-left: 70px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part906"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"><span>○</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part906" data-index="20" style="margin-left: 70px;">PCR program: </div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part907"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part907" data-index="21" style="margin-left: 70px;">98°C/30sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part908"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part908" data-index="22" style="margin-left: 70px;">25 cycles : 	98°C/10sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part909"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 140px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part909" data-index="23" style="margin-left: 140px;">65°C/30sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part910"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 140px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part910" data-index="24" style="margin-left: 140px;">72°C/30sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part911"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part911" data-index="25" style="margin-left: 70px;">72°C/2min</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part912"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part912" data-index="26" style="margin-left: 70px;">infinite 15°C</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part913"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part913" data-index="27" style="margin-left: 70px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part914"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"><span>○</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part914" data-index="28" style="margin-left: 70px;">agarose 1% gel, 100V 25min. Results :</div></div></div><div class="mediocre-item" data-scroll-id="note-part915"><div class=""><div><div class="hiddenFocusEditable editable" contenteditable="true" data-cid="note-part915-item" data-formattable="true" data-index="29" data-subindex="item"></div><div class="mediocre-captionEditable multi-editable"><div class="mediocre-captionEditable-indent" style="padding-left: 35px;"><div class="mediocre-captionEditable-grid"><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell mediocre-captionEditable-component"><div class="note-item-wrapper"><div class="dnd-drop-container"><div class="noteLinkedFilePreview mediocre-itemSelect note-item"><div><h5 class="u-no-spacing-top u-no-spacing-bottom"><div class="pull-right"></div><div class="pull-right"><span></span></div><span class="glyphicon glyphicon-paperclip note-icon u-small-spacing-right pull-left" icon="paperclip"></span><div class="note-itemName">image.png</div></h5><div class="note-preview-body"><img alt="thumbnail" class="noteLinkedFilePreviewImg" src="assets/Qnnrt5BKqHBr8hA9w8Z0c02f4KD3IHbI7XVxvM3O-image.png"/></div></div></div><div class="dnd-drop-container-border"></div></div></div></div><div class="mediocre-captionEditable-cell" style="width: 0px;"></div><div class="mediocre-captionEditable-cell mediocre-captionEditable-caption editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="true" data-cid="note-part915" data-index="29">
+</div></div><div class="mediocre-captionEditable-cell"></div></div></div></div></div></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part905"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part905" data-index="19" style="margin-left: 70px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part906"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"><span>○</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part906" data-index="20" style="margin-left: 70px;">PCR program: </div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part907"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part907" data-index="21" style="margin-left: 70px;">98°C/30sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part908"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part908" data-index="22" style="margin-left: 70px;">25 cycles : 	98°C/10sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part909"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 140px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part909" data-index="23" style="margin-left: 140px;">65°C/30sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part910"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 140px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part910" data-index="24" style="margin-left: 140px;">72°C/30sec</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part911"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part911" data-index="25" style="margin-left: 70px;">72°C/2min</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part912"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part912" data-index="26" style="margin-left: 70px;">infinite 15°C</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part913"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part913" data-index="27" style="margin-left: 70px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part914"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"><span>○</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part914" data-index="28" style="margin-left: 70px;">agarose 1% gel, 100V 25min. Results :</div>
+<img src="https://2019.igem.org/wiki/images/b/bf/T--Go_Paris-Saclay--Asq.png">
+</div></div><div class="mediocre-item" data-scroll-id="note-part915"><div class=""><div><div class="hiddenFocusEditable editable" contenteditable="true" data-cid="note-part915-item" data-formattable="true" data-index="29" data-subindex="item"></div><div class="mediocre-captionEditable multi-editable"><div class="mediocre-captionEditable-indent" style="padding-left: 35px;"><div class="mediocre-captionEditable-grid"><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell mediocre-captionEditable-component"><div class="note-item-wrapper"><div class="dnd-drop-container"><div class="noteLinkedFilePreview mediocre-itemSelect note-item"><div><h5 class="u-no-spacing-top u-no-spacing-bottom"><div class="pull-right"></div><div class="pull-right"><span></span></div><span class="glyphicon glyphicon-paperclip note-icon u-small-spacing-right pull-left" icon="paperclip"></span><div class="note-itemName">image.png</div></h5><div class="note-preview-body"><img alt="thumbnail" class="noteLinkedFilePreviewImg" src="assets/Qnnrt5BKqHBr8hA9w8Z0c02f4KD3IHbI7XVxvM3O-image.png"/></div></div></div><div class="dnd-drop-container-border"></div></div></div></div><div class="mediocre-captionEditable-cell" style="width: 0px;"></div><div class="mediocre-captionEditable-cell mediocre-captionEditable-caption editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="true" data-cid="note-part915" data-index="29">
 </div></div></div></div></div></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part917"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 70px;"><span>○</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part917" data-index="30" style="margin-left: 70px;"><span style="color:#344043">Heatshock plasmid transformation (Natacha)</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part918"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part918" data-index="31" style="margin-left: 35px;"><span style="color:#344043">Let DH5alpha on ice for 30 min</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part919"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part919" data-index="32" style="margin-left: 35px;"><span style="color:#344043">Add plasmid PZA34 and PZA31M2N  (a few µL) in your bacterial solution and let it on ice for 30 min</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part920"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part920" data-index="33" style="margin-left: 35px;">Heatshock at 42°C for 1 min</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part921"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part921" data-index="34" style="margin-left: 35px;"><span style="color:#344043">Put the solution back in ice for 2 min</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part922"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part922" data-index="35" style="margin-left: 35px;"><span style="color:#344043">Add 1 mL LB  (+ Glu) and put the solution at 37°C for 1 h with shaking</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part923"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part923" data-index="36" style="margin-left: 35px;"><span style="color:#344043">Spread out transformants on agar plates (50 µL, 100 µL and 100 µL concentrate)</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part924"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part924" data-index="37" style="margin-left: 35px;"><span style="color:#344043">Put the plates at 37°C overnight</span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part925"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part925" data-index="38" style="margin-left: 0px;"></div></div></div><div class="daySeparator" data-scroll-id="entryDay844"><div class="daySeparator-inputWrapper"><input class="daySeparator-inputWrapper-hiddenInput" type="input"/></div><span class="daySeparator-date">Friday, 12/7/2019</span><div class="daySeparator-line"></div></div><div class="mediocre-item is-text" data-scroll-id="note-part926"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"><span>●</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part926" data-index="39" style="margin-left: 35px;"><b>Plasmid extraction of pZA31, pZA34 and pSB1C3 : Arnaud</b></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part927"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part927" data-index="40" style="margin-left: 0px;">Use of <i>Macherey-Nagel kit "DNA and RNA purification" protocol page 18 "Low copy"</i></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part928"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part928" data-index="41" style="margin-left: 0px;">3 extractions per plasmids pZA31 and pZA34 and 2 extractions for pSB1C3</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part929"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part929" data-index="42" style="margin-left: 0px;">Analysis with Nanodrop :</div></div></div><div class="mediocre-item is-captionedTable" data-scale-for-print="print1907" data-scroll-id="note-part930"><div class=""><div><div class="hiddenFocusEditable editable" contenteditable="true" data-cid="note-part930-item" data-formattable="true" data-index="43" data-subindex="item"></div><div class="mediocre-captionEditable multi-editable mediocre-captionEditable--table"><div class="mediocre-captionEditable-indent"><div class="mediocre-captionEditable-grid"><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell"></div><div class="mediocre-captionEditable-cell mediocre-captionEditable-component" style="width: 100%;"><div class="note-item-wrapper"><div class="mediocre-tableEditable printable-container-scaleForPrint" data-formattable="true" data-index="43" data-printable-layout-width="465" data-subindex="cell"><div></div><div class="hiddenFocusEditable js-focusBearer" contenteditable="true" tabindex="-1"></div><div class="hiddenFocusEditable editable hiddenFocusEditable--fullTable" contenteditable="true" data-cid="note-part930-table" data-formattable="true" data-index="43" data-subindex="table"></div><div class="u-position-relative mediocre-tableEditable-scrollAreaWrapper" style="margin-left: 0px; margin-right: 0px;"><div class="mediocre-tableEditable-scrollArea" style="padding-top: 47px;"><div class="mediocre-tableEditable-tableWrapper" style="padding-left: 0px;"><div class="mediocre-tableEditable-border multi-editable" style="left: -6px; top: -43px; bottom: 4px; width: 457px; min-width: 12px;"></div><div class="u-position-relative u-flex-display"><table class="mediocre-tableEditable-table mediocre-tableEditable-table--main" style="width: 445px;"><colgroup><col style="width: 45px;"></col><col data-id="0" style="width: 100px;"></col><col data-id="1" style="width: 100px;"></col><col data-id="2" style="width: 100px;"></col><col data-id="3" style="width: 100px;"></col></colgroup><tbody><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--header"></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">A</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">A</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">B</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">B</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">C</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">C</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">D</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">D</span><div class="u-flex-display"></div></div></div></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">1</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">1</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-0-0" data-namespace="table-ns1831"><span>Plasmid</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-0-1" data-namespace="table-ns1831"><span>ng/uL</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-0-2" data-namespace="table-ns1831"><span>260/230</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-0-3" data-namespace="table-ns1831"><span>260/280</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">2</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">2</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-1-0" data-namespace="table-ns1831"><span>pSB1C3 (1)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-1-1" data-namespace="table-ns1831"><span>52.4</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-1-2" data-namespace="table-ns1831"><span>2.39</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-1-3" data-namespace="table-ns1831"><span>2.04</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">3</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">3</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-2-0" data-namespace="table-ns1831"><span>pSB1C3 (2)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-2-1" data-namespace="table-ns1831"><span>66.84</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-2-2" data-namespace="table-ns1831"><span>2.21</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-2-3" data-namespace="table-ns1831"><span>2.01</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">4</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">4</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-3-0" data-namespace="table-ns1831"><span>pZA34 (1)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-3-1" data-namespace="table-ns1831"><span>10.57</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-3-2" data-namespace="table-ns1831"><span>1.09</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-3-3" data-namespace="table-ns1831"><span>2.56</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">5</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">5</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-4-0" data-namespace="table-ns1831"><span>pZA34 (2)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-4-1" data-namespace="table-ns1831"><span>13.08</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-4-2" data-namespace="table-ns1831"><span>1.15</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-4-3" data-namespace="table-ns1831"><span>2.21</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">6</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">6</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-5-0" data-namespace="table-ns1831"><span>pZA34 (3)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-5-1" data-namespace="table-ns1831"><span>12.34</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-5-2" data-namespace="table-ns1831"><span>1.61</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-5-3" data-namespace="table-ns1831"><span>2.51</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">7</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">7</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-6-0" data-namespace="table-ns1831"><span>pZA31 (1)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-6-1" data-namespace="table-ns1831"><span>12.74</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-6-2" data-namespace="table-ns1831"><span>2.10</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-6-3" data-namespace="table-ns1831"><span>2.70</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">8</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">8</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-7-0" data-namespace="table-ns1831"><span>pZA31 (2)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-7-1" data-namespace="table-ns1831"><span>14.35</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-7-2" data-namespace="table-ns1831"><span>1.40</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-7-3" data-namespace="table-ns1831"><span>2.63</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">9</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">9</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part930-8-0" data-namespace="table-ns1831"><span>pZA31 (3)</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-8-1" data-namespace="table-ns1831"><span>13.79</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-8-2" data-namespace="table-ns1831"><span>2.08</span></td><td class="cell is-value is-alignRight cell-contents" data-cid="table-note-part930-8-3" data-namespace="table-ns1831"><span>2.61</span></td></tr></tbody><tfoot></tfoot></table><div class="mediocre-tableEditable-fillerTableWrapper" style="margin-right: 10px;"><table class="mediocre-tableEditable-fillerTable" style="display: none;"><tbody><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr><tr style="height: 0px;"><td></td></tr></tbody></table></div></div></div></div></div><div class="mediocre-tableEditable-titleBarWrapper"><div class="mediocre-tableEditable-titleBar" style="height: 37px; padding-left: 0px;"><div class="mediocre-tableEditable-tableName u-pointer"><div class="u-flex-grow u-overflow-hidden u-verticalCenterItems"><div class="u-ellipsis-text"><span class="u-ellipsis-text">Table4</span></div></div><div class="u-flex-none"></div></div><div class="u-flex-none u-tiny-spacing-left u-verticalCenterItems table-floater"><svg class="svgIcon workflowTable-tableCollapser u-pointer"><use xlink:href="#chevron-up_c0f0f77c91b0d9072b0d8255f9f54e3c"></use></svg></div></div></div></div></div></div><div class="mediocre-captionEditable-cell"></div></div><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell"></div><div class="mediocre-captionEditable-cell"><div class="mediocre-captionEditable-caption--bottom mediocre-captionEditable-caption editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="true" data-cid="note-part930" data-index="43" style="max-width: 465px; margin-left: 0px;">
 </div></div><div class="mediocre-captionEditable-cell"></div></div></div></div></div></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part931"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 35px;"><span>●</span></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part931" data-index="44" style="margin-left: 35px;"><b>PCR for Gibson assembly of pBAD24A1/pBAD24_Moclo: Kim</b></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part932"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part932" data-index="45" style="margin-left: 0px;">PCR of plasmid <span style="color:#344043">pBAD24 with or without the A1 gene. </span></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part933"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part933" data-index="46" style="margin-left: 0px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part934"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part934" data-index="47" style="margin-left: 0px;">Expected size : pBAD24_A1 = 5,5 kB</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part935"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part935" data-index="48" style="margin-left: 0px;">                            pBAD24_Moclo = 3,9 kB</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part936"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part936" data-index="49" style="margin-left: 0px;">                            A1 = 1,6 kB</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part937"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part937" data-index="50" style="margin-left: 0px;"></div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part938"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part938" data-index="51" style="margin-left: 0px;"><b>Primers</b>: pBAD24_F (9) and pBAD24_R (10)</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part939"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part939" data-index="52" style="margin-left: 0px;">                Tm = 62°C</div></div></div><div class="mediocre-item is-text" data-scroll-id="note-part940"><div class="mediocre-noteEditable multi-editable is-paragraph"><div class="mediocre-noteEditable-indenter" style="width: 0px;"></div><div class="editable selectable is-locked" contenteditable="false" data-allow-at-search="true" data-allow-multiline="false" data-cid="note-part940" data-index="53" style="margin-left: 0px;"></div></div></div><div class="mediocre-item is-captionedTable" data-scale-for-print="print1908" data-scroll-id="note-part941"><div class=""><div><div class="hiddenFocusEditable editable" contenteditable="true" data-cid="note-part941-item" data-formattable="true" data-index="54" data-subindex="item"></div><div class="mediocre-captionEditable multi-editable mediocre-captionEditable--table"><div class="mediocre-captionEditable-indent"><div class="mediocre-captionEditable-grid"><div class="mediocre-captionEditable-row"><div class="mediocre-captionEditable-cell"></div><div class="mediocre-captionEditable-cell mediocre-captionEditable-component" style="width: 100%;"><div class="note-item-wrapper"><div class="mediocre-tableEditable printable-container-scaleForPrint" data-formattable="true" data-index="54" data-printable-layout-width="763" data-subindex="cell"><div></div><div class="hiddenFocusEditable js-focusBearer" contenteditable="true" tabindex="-1"></div><div class="hiddenFocusEditable editable hiddenFocusEditable--fullTable" contenteditable="true" data-cid="note-part941-table" data-formattable="true" data-index="54" data-subindex="table"></div><div class="u-position-relative mediocre-tableEditable-scrollAreaWrapper" style="margin-left: 0px; margin-right: 0px;"><div class="mediocre-tableEditable-scrollArea" style="padding-top: 47px;"><div class="mediocre-tableEditable-tableWrapper" style="padding-left: 0px;"><div class="mediocre-tableEditable-border multi-editable" style="left: -6px; top: -43px; bottom: 4px; width: 755px; min-width: 12px;"></div><div class="u-position-relative u-flex-display"><table class="mediocre-tableEditable-table mediocre-tableEditable-table--main" style="width: 743px;"><colgroup><col style="width: 45px;"></col><col data-id="0" style="width: 207px;"></col><col data-id="1" style="width: 365px;"></col><col data-id="2" style="width: 126px;"></col></colgroup><tbody><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--header"></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">A</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">A</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">B</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">B</span><div class="u-flex-display"></div></div></div></td><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--col"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible mediocre-tableEditable-axisCell--header">C</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text mediocre-tableEditable-axisCell--header">C</span><div class="u-flex-display"></div></div></div></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">1</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">1</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part941-0-0" data-namespace="table-ns1831"><span>Primer name</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part941-0-1" data-namespace="table-ns1831"><span>Primer sequence</span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part941-0-2" data-namespace="table-ns1831"><span>Tm °C</span></td></tr><tr><td class="mediocre-tableEditable-axisCell mediocre-tableEditable-axisCell--row"><div class="mediocre-tableEditable-axisCell-labelWrapper"><div class="mediocre-tableEditable-axisCell-labelSizer u-overflow-wrap-break-word invisible">2</div><div class="u-flex-display invisible"></div><div class="mediocre-tableEditable-axisCell-label u-verticalCenterItems"><span class="mediocre-tableEditable-axisCell-label-text u-ellipsis-text">2</span><div class="u-flex-display"></div></div></div></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part941-1-0" data-namespace="table-ns1831"><span>2019GO-9- ori15A_pBAD24_v2_F  </span></td><td class="cell is-text is-alignLeft cell-contents" data-cid="table-note-part941-1-1" data-namespace="table-ns1831"><span>gtccaagcgagctcgatatcgagcctatggaaaaacgcca

Difference between revisions of "Team:GO Paris-Saclay/Cleaningfactory"

Revision as of 16:30, 20 October 2019

Cleaning factory: methotrexate

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Gel PCR 15.07.jpg

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PCR pbadmoclo et pbad A1 T7.pdf