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<div class="submenu"> | <div class="submenu"> | ||
<a href="/Team:Humboldt_Berlin/Team">Team members</a> | <a href="/Team:Humboldt_Berlin/Team">Team members</a> | ||
− | <a href="/Team:Humboldt_Berlin/Collaborations"> | + | <a href="/Team:Humboldt_Berlin/Collaborations">Collaborations</a> |
</div> | </div> | ||
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<a href="/Team:Humboldt_Berlin/Description">Description</a> | <a href="/Team:Humboldt_Berlin/Description">Description</a> | ||
<a href="/Team:Humboldt_Berlin/Design">Design</a> | <a href="/Team:Humboldt_Berlin/Design">Design</a> | ||
− | <a href="/Team:Humboldt_Berlin/Experiments"> | + | <a href="/Team:Humboldt_Berlin/Experiments">Experiments</a> |
<a class="active" href="/Team:Humboldt_Berlin/Notebook">Notebook</a> | <a class="active" href="/Team:Humboldt_Berlin/Notebook">Notebook</a> | ||
<a href="/Team:Humboldt_Berlin/Contribution">Contribution</a> | <a href="/Team:Humboldt_Berlin/Contribution">Contribution</a> | ||
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<a href="/Team:Humboldt_Berlin/Human_Practices">Human Practices</a> | <a href="/Team:Humboldt_Berlin/Human_Practices">Human Practices</a> | ||
<div class="submenu"> | <div class="submenu"> | ||
− | <a href="/Team:Humboldt_Berlin/Human_Practices">Human | + | <a href="/Team:Humboldt_Berlin/Human_Practices">Human Practices</a> |
<a href="/Team:Humboldt_Berlin/Public_Engagement">Education & Engagement</a> | <a href="/Team:Humboldt_Berlin/Public_Engagement">Education & Engagement</a> | ||
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<a>Awards</a> | <a>Awards</a> | ||
<div class="submenu"> | <div class="submenu"> | ||
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<a href="/Team:Humboldt_Berlin/Hardware">Hardware</a> | <a href="/Team:Humboldt_Berlin/Hardware">Hardware</a> | ||
<a href="/Team:Humboldt_Berlin/Measurement">Measurement</a> | <a href="/Team:Humboldt_Berlin/Measurement">Measurement</a> | ||
<a href="/Team:Humboldt_Berlin/Model">Model</a> | <a href="/Team:Humboldt_Berlin/Model">Model</a> | ||
<a href="/Team:Humboldt_Berlin/Plant">Plant</a> | <a href="/Team:Humboldt_Berlin/Plant">Plant</a> | ||
− | |||
</div> | </div> | ||
</div> | </div> | ||
<div class="devider"></div> | <div class="devider"></div> | ||
− | <a href="https://igem.org/ | + | <a href="https://2019.igem.org/Team:Humboldt_Berlin/Achievements"> |
For Judges | For Judges | ||
</a> | </a> | ||
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</nav> | </nav> | ||
− | < | + | <div class="fixed-header-container"> |
− | + | <section class="fixed-image-header"> | |
− | + | <img src="https://static.igem.org/mediawiki/2019/8/81/T--Humboldt_Berlin--Notebook_Header.jpeg" alt="notebook" /> | |
− | + | </section> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | < | + | <h1 class="page-headline description">Notebook</h1> |
− | + | </div> | |
− | + | ||
− | + | <div class="full-height fixed-header-content notebook"> | |
− | + | <div class="width-limit padding-container" style="margin: 50px 0;"> | |
− | [].forEach.call( | + | <img src="https://static.igem.org/mediawiki/2019/e/e3/T--Humboldt_Berlin--notebook_final.png" alt="our stuffed calendar"/></br> |
− | + | </div> | |
− | el. | + | <section class="width-limit"> |
− | } | + | <div class="category-container"> |
− | + | <h3>Select category</h3> | |
− | + | <div class="category" data-category="synthesis"> | |
+ | <img src="https://static.igem.org/mediawiki/2019/6/6f/T--Humboldt_Berlin--label_notebook_synthesis.png" alt="Synthesis: Parts and Vectors" /> | ||
+ | </div> | ||
+ | <div class="category" data-category="human-practice"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/e/e6/T--Humboldt_Berlin--label_notebook_human_practise.png" alt="human practice" /> | ||
+ | </div> | ||
+ | <div class="category" data-category="chlamy"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/a/ac/T--Humboldt_Berlin--label_notebook_chlamy.png" alt="c. reinhardtii cultivation and transformations"/> | ||
+ | </div> | ||
+ | <!--<div class="category" data-category="bioreaktor"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/6/66/T--Humboldt_Berlin--label_notebook_bioreaktor.png" alt="bioreaktor" /> | ||
+ | </div>--> | ||
+ | </div> | ||
+ | <div class="timeline"> | ||
+ | <!-- synthesis --> | ||
+ | <div class="timeline-month tl-first-element" | ||
+ | data-text="- Learning MoClo/Golden Gate Design <br> | ||
+ | - Research on PETase degradation <br> | ||
+ | - Planning construction design for Golden Gate cloning using 10 different fusion sites according to Patron <br> | ||
+ | - Designing Primers for <i>C. reinhardtii</i> specified promotor PsaD containing BpiI recognition site with MoClo fusion sites and fusion sites compatible for L0 backbone. Primers containing mutation to delete BpiI site (Primer 5,6,11-13)." | ||
+ | data-date="10/08 - 10/14" | ||
+ | data-category="synthesis"> | ||
+ | October <br> 2018 | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="- Research on <i>C. reinhardtii</i> cultivation <br> | ||
+ | <h2>YFP</h2> | ||
+ | - Designing Primers for YFP with B5-B5 fusion sites taken from AG Hegemann plasmid p135 (Primer 7+8)" | ||
+ | data-date="10/15 - 10/21" | ||
+ | data-category="synthesis"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="- Searching for gene templates <br> | ||
+ | <h2>chemical competent DH10B</h2> | ||
+ | - production of chemical competent E. coli DH10B cells <br> | ||
+ | - PCR for YFP B5-B5 with primer 7+8" | ||
+ | data-category="synthesis" | ||
+ | data-date="10/22 - 10/28" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/e/eb/T--Humboldt_Berlin--notebook_s_1022.png"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>chemical competent DH10B</h2> | ||
+ | - checking the chemical competent DH10B cells → Transformation of plasmid containing RFP (Plasmid: pP51) (result/picture: red colonies grew → cells are competent! no cells on negative control) | ||
+ | <h2>ARS</h2> | ||
+ | - Design of Arylsulfatase 1 (ARS) secretion signal as a B2-B2 part | ||
+ | <h2>GLE</h2> | ||
+ | - Design of Gametolysin (GLE) secretion signal as a B2-B2 part" | ||
+ | data-date="10/29 - 11/04" | ||
+ | data-category="synthesis" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/1/17/T--Humboldt_Berlin--Notebook-10-29.png"> | ||
+ | November | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>PsaD</h2> | ||
+ | - 11+12 → fragment 1 <br> | ||
+ | - 12+6 → fragment 2 <br> | ||
+ | - 13+5 → fragment 3 <br> | ||
+ | → result: PCR successful <br> | ||
+ | - Second PRC to combine PsaD fragments: <br> | ||
+ | fragment 2 + 3 → part 1 <br> | ||
+ | fragment 1 + 3 → part 2 <br> | ||
+ | → result: PCR successful" | ||
+ | data-category="synthesis" | ||
+ | data-date="11/05 - 11/11" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/9/9f/T--Humboldt_Berlin--notebook_s_115.png"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<div class='two-columns'><div> | ||
+ | <h2>PsaD</h2> | ||
+ | - Third PRC to amplify PsaD from outer sites <br> | ||
+ | 1) Template: part 1; primer: 5+6<br> | ||
+ | 2) Template: Part 2; Primer: 5+6<br> | ||
+ | result: PCR was successful <br> | ||
+ | Clean-Up of PCR products <br> | ||
+ | Golden Gate Restriction/Ligation of PsaD PCR Products (PsaD A1-A3 and PsaD A1-B1) with Level 0 plasmid using <i>Bpi</i>I | ||
+ | <h2>YFP</h2> | ||
+ | Recognition of frameshift in YFP; new design of Primers for p51 from AG Hegemann <br> | ||
+ | - Primer 24+25 → YFP B5-B5 <br> | ||
+ | - Primer 26 + 27 → YFP B3-B4 <br> | ||
+ | <h2>RFP</h2> | ||
+ | - Designing Primers to amplify RFP containing <i>Bpi</i>I and <i>Bsa</i>I recognition site (Primer 39/40) to build a modulated L0-backbone | ||
+ | </div> | ||
+ | <div> | ||
+ | <h2>ARS1</h2> | ||
+ | - New design of ARS1 secretion signal as a B2-B2 part <br> | ||
+ | - Oligos 16 + 17 were ordered | ||
+ | <h2>GLE</h2> | ||
+ | - New design of GLE secretion signal as a B2-B2 part <br> | ||
+ | - Oligos 18 + 19 were ordered | ||
+ | <h2>SP20</h2> | ||
+ | -Design of SP20 secretion enhancement as a B5-B5 part<br> | ||
+ | - Oligos 20 + 21 were ordered | ||
+ | </div></div>" | ||
+ | data-category="synthesis" | ||
+ | data-date="11/12 - 11/18" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/8/8c/T--Humboldt_Berlin--notebook_s_1112_2.png"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>RFP</h2> | ||
+ | - Designing Primers to amplify RFP containing <i>Bpi</i>I and <i>Bsa</i>I recognition site (Primer 39/40) to build a modulated L0-backbone | ||
+ | <h2>PsaD</h2> | ||
+ | - Heatshock Transformation of L0-PsaD A1/A3 and L0-PsaD A1/B1 into heat-competent <i>E.coli</i> DH10B | ||
+ | Result/Picture: white colonies carrying L0-PsaD, blue colonies just L0 | ||
+ | YFP Golden Gate Restriction/Ligation of YFP B5-B5 and YFP B3-B4 into L0" | ||
+ | data-category="synthesis" | ||
+ | data-date="11/12 - 11/18" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/9/93/T--Humboldt_Berlin--notebook_s_1119.png"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>PsaD</h2> | ||
+ | - overnight cultures of DH10B with L0-PsaD A1/A3 and L0-PsaD A1/B1 | ||
+ | <div style='text-indent:20px;'>- plasmid prep of ON-culture</div> | ||
+ | <div style='text-indent:20px;'>- Nano-Drop concentration</div> | ||
+ | - sequencing result: positive sequencing result of 1 clone each" | ||
+ | data-date="11/26 - 12/02" | ||
+ | data-category="synthesis"> | ||
+ | December | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<div class='two-columns'><div> | ||
+ | <h2>RFP</h2> | ||
+ | - PCR to amplify RFP from pPH041 backbone using primers 39/40 <br> | ||
+ | - RFP will be selection marker on L0 backbone within the fusion site <br> | ||
+ | - restriction of RFP with <i>Bsa</i>I; restriction of pAGM9121 backbone with <i>Bsa</i>I <br> | ||
+ | - Ligation of pAGM9121 backbone with RFP using T4 ligase → L0-RFP (RFP ist NOT an insert, its inside the cloning site) <br> | ||
+ | - Heatshock transformation of L0-RFP into heat-competent <i>E.coli</i> DH10B | ||
+ | </div> | ||
+ | <div> | ||
+ | <h2>ARS1</h2> | ||
+ | - Oligoannealing of Oligos 16 + 17 for the creation of the ARS1 secretion signal with B2 fusion sites | ||
+ | <h2>GLE</h2> | ||
+ | - Oligoannealing of Oligos 18 + 19 for the creation of the GLE secretion signal with B2 fusion sites | ||
+ | <h2>SP20</h2> | ||
+ | - Oligoannealing of Oligos 20 + 21 for the creation of the SP20 secretion enhancement with B5 fusion sites | ||
+ | </div></div>" | ||
+ | data-category="synthesis" | ||
+ | data-date="12/03 - 12/09" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/3/3a/T--Humboldt_Berlin--notebook_s_1203.png"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<div class='two-columns'><div> | ||
+ | <h2>L0-RFP</h2> | ||
+ | - positive sequencing result of L0-RFP containing required restriction and fusion sites | ||
+ | <h2>PsaD</h2> | ||
+ | - Golden Gate Restriction/Ligation of PsaD PCR Products (PsaD A1-A3 and PsaD A1-B1) with L0-RFP plasmid using <i>Bpi</i>I | ||
+ | <h2>ARS1</h2> | ||
+ | - Ligation of ARS1 into L0-RFP<br> | ||
+ | - Heatshock transformation of L0-ARS into heat-competent <i>E.coli</i> DH10B<br> | ||
+ | - 3 Over-night cultures were made and prepped and sent for sequencing | ||
+ | <h2>GLE</h2> | ||
+ | - Ligation of GLE into L0-RFP<br> | ||
+ | - Heatshock transformation of L0-GLE into heat-competent <i>E.coli</i> DH10B<br> | ||
+ | - 3 Over-night cultures were made and prepped and sent for sequencing | ||
+ | </div> | ||
+ | <div> | ||
+ | <h2>SP20</h2> | ||
+ | - Ligation of SP20 into L0-RFP<br> | ||
+ | - Heatshock transformation of L0-SP20 into heat-competent <i>E.coli</i> DH10B<br> | ||
+ | - 3 Over-night cultures were made and prepped and sent for sequencing | ||
+ | <h2>His-Tag</h2> | ||
+ | - Design of a 6xHis-Tag for detection, isolation and purification as a C-terminal tag; B5-B5 module<br> | ||
+ | - Oligonucleotides 47+48 were ordered | ||
+ | <h2>HA-Tag</h2> | ||
+ | - Design of a 3xHA-Tag for detection, isolation and purification as a C-terminal tag; B5-B5 module<br> | ||
+ | - Oligonucleotides 49+50 were ordered | ||
+ | </div></div>" | ||
+ | data-category="synthesis" | ||
+ | data-date="12/10 - 12/16" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/e/e1/T--Humboldt_Berlin--notebook_s_1210_1.png"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<div class='two-columns'><div> | ||
+ | <h2>YFP</h2> | ||
+ | - Heatshock Transformation of L0-YFP B5-B5 and B3-B4 into heat-competent <i>E.coli</i> DH10B | ||
+ | 3 Over-night cultures were made and prepped and sent for sequencing | ||
+ | <h2>PsaD</h2> | ||
+ | - Heatshock Transformation of L0-PsaD A1/A3 and L0-PsaD A1/B1 into heat-competent <i>E.coli</i> DH10B <br> | ||
+ | - Result/Picture: white colonies carrying L0-PsaD, red colonies just L0-RFP | ||
+ | </div> | ||
+ | <div> | ||
+ | <h2>His-Tag</h2> | ||
+ | - Oligoannealing of 47+48 to produce the 6xHis-Tag with B5-B5 fusion sites<br> | ||
+ | - Ligation of the annealed product to L0-RFP | ||
+ | <h2>HA-Tag</h2> | ||
+ | - Oligoannealing of 49+50 to produce the 3xHA-Tag with B5-B5 fusion sites<br> | ||
+ | - Ligation of the annealed product to L0-RFP | ||
+ | </div></div>" | ||
+ | data-category="synthesis" | ||
+ | data-date="12/17 - 12/23" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/7/77/T--Humboldt_Berlin--notebook_s_1217_1.png"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>YFP</h2> | ||
+ | - Positive sequencing results of L0-YFP B5-B5 and B3-B4 | ||
+ | <h2>ARS1</h2> | ||
+ | - Positive sequencing results of L0-ARS B2-B2 | ||
+ | <h2>GLE</h2> | ||
+ | - Positive sequencing results of L0-GLE B2-B2 | ||
+ | <h2>SP20</h2> | ||
+ | - Positive sequencing results of L0-SP20 B5-B5 | ||
+ | <h2>PsaD</h2> | ||
+ | - Positive sequencing results of L0-PsaD A1/A3 <br> | ||
+ | - negative sequencing results of L0-PsaD A1/B1" | ||
+ | data-date="12/31 - 01/06" | ||
+ | data-category="synthesis"> | ||
+ | January | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>His-Tag</h2> | ||
+ | - Failed Heatshock Transformation of L0-His-Tag into heat-competent <i>E.coli</i> DH10B (07.01.) <br> | ||
+ | - Repetition of Heatshock transformation (10.01.) <br> | ||
+ | - 3 Over-night cultures were made and prepped and sent for sequencing <br> | ||
+ | <h2>HA-Tag</h2> | ||
+ | Failed Heatshock Transformation of L0-HA-Tag into heat-competent <i>E.coli</i> DH10B (07.01.) <br> | ||
+ | Repetition of Heatshock transformation (10.01) <br> | ||
+ | 3 Over-night cultures were made and prepped and sent for sequencing" | ||
+ | data-category="synthesis" | ||
+ | data-date="01/07 - 01/13" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/5/5b/T--Humboldt_Berlin--notebook_s_1217_4.png"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>His-Tag</h2> | ||
+ | - Oligoannealing of 47+55 to produce the 6xHis-Tag with B5-B5 fusion sites <br> | ||
+ | - Ligation of the annealed product to L0-RFP <br> | ||
+ | - Heatshock Transformation of L0-His-Tag into heat-competent <i>E.coli</i> DH10B <br> | ||
+ | - Colony PCR <br> | ||
+ | - 3 Over-night cultures were made and prepped and sent for sequencing" | ||
+ | data-date="01/28 - 02/03" | ||
+ | data-category="synthesis" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/c/ca/T--Humboldt_Berlin--notebook_s_0128_1.png"> | ||
+ | February | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>His-Tag</h2> | ||
+ | - Corrected sequencing results of L0-6xHis-Tag B5-B5 | ||
+ | <h2>B2 Linker for Level 1 plasmid</h2> | ||
+ | - Designing DNA Oligos as a Linker module containing B2 fusion sites ('Primer' 72/73)" | ||
+ | data-category="synthesis" | ||
+ | data-date="02/04 - 02/10"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>Paromomycin</h2> | ||
+ | - Designing primers for aphVIII with B3-B4 fusion sites taken from AG Hegemann plasmid p114 (Primer 75+76) | ||
+ | <h2>Hygromycin</h2> | ||
+ | - Designing primers for aphVII with B3-B4 fusion sites taken from AG Hegemann plasmid p360 (Primer 77+78) | ||
+ | <h2>B2 Linker for Level 1 plasmid</h2> | ||
+ | - Oligo Annealing of B2 Linker using Primers 72/73" | ||
+ | data-category="synthesis" | ||
+ | data-date="02/11 - 02/17"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<div class='two-columns'><div> | ||
+ | <h2>Paromomycin</h2> | ||
+ | - <b>02/18</b> Doing a PCR with primers 75+76 and p114; PCR-clean up<br> | ||
+ | - <b>02/20</b> Golden Gate Restriction/Ligation of Paromomycin PCR Product (Paro B3-B4) with L0-RFP plasmid using <i>Bpi</i>I<br> | ||
+ | - <b>02/22</b> Heatshock Transformation of L0-Paro B3-B4 into heat-competent <i>E.coli</i> DH10B<br> | ||
+ | - Colony PCR | ||
+ | <h2>Hygromycin</h2> | ||
+ | - <b>02/18</b> Doing a PCR with primers 77+78 and p360 → Failed<br> | ||
+ | - <b>02/19</b> Repeating PCR with primers 77+78 and p360 → Failed<br> | ||
+ | - Optimization of annealing temperature to 62°C → we got the wanted product<br> | ||
+ | - <b>02/21</b> Golden Gate Restriction/Ligation of Hygromycin PCR Product (Hyg B3-B4) with L0-RFP plasmid using <i>Bpi</i>I<br> | ||
+ | - <b>02/22</b> Heatshock Transformation of L0-Hyg B3-B4 into heat-competent <i>E.coli</i> DH10B<br> | ||
+ | - Colony PCR with white colonies → Failed | ||
+ | </div> | ||
+ | <div> | ||
+ | <h2>Amp/Ori part for level 1 plasmid</h2> | ||
+ | - PCR to amplify Amp/Ori from pICH47732 backbone using primers 56/57<br> | ||
+ | - restriction of Amp/Ori with <i>Xba</i>I and BamHI | ||
+ | <h2>L1a,b,c-RFP part for level 1 plasmid</h2> | ||
+ | - PCR to amplify RFP from L0-RFP backbone using different primers to get 3 different L1 overhang versions<br> | ||
+ | -- L1a-RFP: Primer 58,59<br> | ||
+ | -- L1b-RFP: Primer: 60, 61<br> | ||
+ | -- L1c-RFP: Primer: 62, 63<br> | ||
+ | - Digestion of different PCR products using <i>Spe</i>I & <i>BamH</i>I | ||
+ | <h2>L1 Plasmids</h2> | ||
+ | - Ligation of Amp/Ori with<br> | ||
+ | -- 1) L1a-RFP<br> | ||
+ | -- 2) L1b-RFP<br> | ||
+ | -- 3) L1c-RFP using T4 ligase<br> | ||
+ | - Heat-shock Transformation into competent DH10B cells Result: not succesful | ||
+ | </div></div>" | ||
+ | data-category="synthesis" | ||
+ | data-date="02/18 - 02/24" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/b/be/T--Humboldt_Berlin--notebook_s_0218_1.png"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<div class='two-columns'><div> | ||
+ | <h2>Paromomycin</h2> | ||
+ | - Preparation of 3 overnight cultures of clones 1, 2, 3 (colony PCR 22.02.) <br> | ||
+ | - Plasmid prep the next day<br> | ||
+ | - Sending plasmids for sequencing <br> | ||
+ | - Corrected sequencing results of L0-Paro B3-B4 | ||
+ | <h2>Hygromycin</h2> | ||
+ | - <b>03/26</b> Second try of doing a colony PCR → worked out<br> | ||
+ | - Preparation of 3 overnight cultures K4,7,9<br> | ||
+ | - <b>03/27</b> plasmid prep + sending them in for sequencing<br> | ||
+ | - <b>03/28</b> Corrected sequencing results of L0-Hyg B3-B4 | ||
+ | </div> | ||
+ | <div> | ||
+ | <h2>L1a,b,c - RFP</h2> | ||
+ | - mistake: it was noticed that primer to amplify 3 different RFP versions were designed wrong → buffer bases were missing, so that the restriction enzymes did not attach to the DNA <br> | ||
+ | - designing and ordering new primers containing buffer bases (primer 84-89) | ||
+ | </div></div>" | ||
+ | data-date="02/25 - 03/03" | ||
+ | data-category="synthesis" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/b/bf/T--Humboldt_Berlin--notebook_s_0225_1.png"> | ||
+ | March | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>L1a,c,b, -RFP</h2> | ||
+ | - new PCR to amplify different RFP versions" | ||
+ | data-category="synthesis" | ||
+ | data-date="03/11 - 03/17"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>YFP</h2> | ||
+ | - Primer design for a new YFP B4-B4 contruct (Primer 111 + 27)" | ||
+ | data-category="synthesis" | ||
+ | data-date="05/13 - 05/19"> | ||
+ | May | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>YFP</h2> | ||
+ | - PCR of YFP B4-B4; size: 749 bp <br> | ||
+ | - Ligation YFP B4-B4 with L0-RFP" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/f/f7/T--Humboldt_Berlin--notebook_s_0527.png" | ||
+ | data-category="synthesis" | ||
+ | data-date="05/27 - 06/02"> | ||
+ | June | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>YFP</h2> | ||
+ | - Transformation L0-YFP B4-B4 into <i>E. coli </i><br> | ||
+ | - Colony PCR YFP-B4-B4 in <i>E. coli</i>" | ||
+ | data-category="synthesis" | ||
+ | data-date="06/03 - 06/09"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>YFP</h2> | ||
+ | - sequencing result of YFP B4-B4 clone 2 was positive" | ||
+ | data-category="synthesis" | ||
+ | data-date="06/17 - 06/23"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>B2 Linker</h2> | ||
+ | - creating new primers" | ||
+ | data-category="synthesis" | ||
+ | data-date="07/01 - 07/07"> | ||
+ | July | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>B2 Linker</h2> | ||
+ | - Oligo Annealing B2 Linker<br> | ||
+ | - succesfull" | ||
+ | data-category="synthesis" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/e/e8/T--Humboldt_Berlin--notebook_s_0708.png" | ||
+ | data-date="06/17 - 06/23"> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <!-- human practice --> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>Green Week Berlin/ Microplastic presentation of BfR</h2> | ||
+ | - participants: Fabienne, Andrej" | ||
+ | data-date="January 2019" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/4/45/T--Humboldt_Berlin--notebook_h_Jan19.jpeg" | ||
+ | data-category="human-practice"> | ||
+ | January 2019 | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>TV report about “Chlamylicious”</h2> | ||
+ | - participants: Fabienne, Juli, Andrej, Dimitri" | ||
+ | data-date="May 2019" | ||
+ | data-category="human-practice" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/2/24/T--Humboldt_Berlin--notebook_h_May19.png"> | ||
+ | May | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>2 week study at MPI Göttingen</h2> | ||
+ | - participant: Darius" | ||
+ | data-category="human-practice" | ||
+ | data-date="May 2019"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>Upcycling Project at the family event of Humboldt University with the collaboration of Satch backpacks</h2> | ||
+ | - participants: Sophia, Luise, Fabienne" | ||
+ | data-category="human-practice" | ||
+ | data-date="May 2019" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/f/fa/T--Humboldt_Berlin--notebook_h_May19_2.png"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>Long Night of The Sciences Berlin</h2> | ||
+ | - participants: Juli, Darius, Luise, Paul, Johannes, Andrej, Ale, Patrick, Marc, Elena, Dimitri, Sandra, Fabienne, Saskia" | ||
+ | data-date="June 2019" | ||
+ | data-category="human-practice" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/8/8b/T--Humboldt_Berlin--notebook_h_Jun19.png"> | ||
+ | June | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>Visit at MINT Microalgae Engineering</h2> | ||
+ | - participants: Paul, Darius, Dimitri, Andrej" | ||
+ | data-date="July 2019" | ||
+ | data-category="human-practice" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/6/69/T--Humboldt_Berlin--notebook_h_Jul19_1.png"> | ||
+ | July | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>Visit and presentation at ResearchGate Berlin</h2> | ||
+ | - participants: Juli, Darius, Andrej, Paul" | ||
+ | data-category="human-practice" | ||
+ | data-date="July 2019" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/2/2b/T--Humboldt_Berlin--notebook_h_Jul19_2.png"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>Visit and project presentation at MERCK</h2> | ||
+ | - participants: Andrej, Dimitri, Darius, Sophia, Saskia, Juli" | ||
+ | data-category="human-practice" | ||
+ | data-date="July 2019" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/5/5a/T--Humboldt_Berlin--notebook_h_Jul19_4.png"> | ||
+ | </div> | ||
+ | <div class="timeline-dot" | ||
+ | data-text="<h2>Visit at algae farm</h2> | ||
+ | - participants: Andrej, Dimitri, Darius, Fabienne, Johannes, Luise, Paul, Simon" | ||
+ | data-category="human-practice" | ||
+ | data-date="July 2019" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/b/b2/T--Humboldt_Berlin--notebook_h_Jul_5.jpeg"> | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>Panel discussion about genetic editing</h2> | ||
+ | - participants: Luise, Sophia, Fabienne" | ||
+ | data-date="October 2019" | ||
+ | data-category="human-practice" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/5/5e/T--Humboldt_Berlin--notebook_h_oct19_3.png"> | ||
+ | October | ||
+ | </div> | ||
+ | |||
+ | <!-- reinhardtii cultivation and transformations (chlamy) --> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>Cultivation of <i>C. reinhardtii</i> in the lab started, introductions to sterile working conditions</h2> | ||
+ | - participants: Alexander Bergmüller, Simon Kelterborn" | ||
+ | data-date="November 2018" | ||
+ | data-category="chlamy"> | ||
+ | November 2018 | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="<h2>First toxicity-tests on algal growth (strain CC-125) of ethylene glycol at concentrations between 0,005-5 % yields no results</h2> | ||
+ | - participants: Alexander Bergmüller" | ||
+ | data-date="November 2018 - December 2018" | ||
+ | data-category="chlamy"> | ||
+ | December | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="- Toxicity-tests: Inhibition of algal growth (strains CC-125 and CC-3403) by ethylene glycol at a concentration between 6-7 % was shown<br> | ||
+ | - Terephthalic acid shown not to inhibit algal growth<br><br> | ||
+ | - participants: Alexander Bergmüller, Sophia Schiebler Darius Rauch, Paul Herrmann" | ||
+ | data-date="January 2019" | ||
+ | data-category="chlamy"> | ||
+ | January | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="- First Toxicity-tests in a multicultivator, which measures algal growth under different conditions: Inhibition of C. reinhardtii by ethylene glycol at a concentration of 6 % was confirmed<br> | ||
+ | - Cultivation of strain CC-125 in high salt medium shows inhibited growth <br><br> | ||
+ | - participants: Paul Herrmann, Andrej Andjelic" | ||
+ | data-date="May 2019" | ||
+ | data-category="chlamy"> | ||
+ | May | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="- started the transformations of L1-constructs into Chlamy<br> | ||
+ | - checked the functionality of our paromomycin and hygromycin selection cassettes<br> | ||
+ | - Cultivation tests of strain SAG 11-32b in high salt medium with added PET in concentrations of 10-800 mg/L. Wastewater contains 250 mg/L PET before treatment. <br><br> | ||
+ | - participants: Simon Kelterborn, Darius Rauch, Dimitri Schumacher, Elena Rätsch, Juliana Rojas Pión, Paul Herrmann" | ||
+ | data-date="June 2019" | ||
+ | data-image="https://static.igem.org/mediawiki/2019/4/40/T--Humboldt_Berlin--HSM_PET.jpeg" | ||
+ | data-category="chlamy"> | ||
+ | June | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="- First Chlamy transformations are discarded, since they were contaminated<br> | ||
+ | - Transformation of further L1-constructs<br> | ||
+ | - constructs containing YFP: colony-PCR showed integration of the plasmid into Chlamy’s genome, but no fluorescence could be measured by the plate reader <br> | ||
+ | - PETase-YFP-containing constructs displayed fluorescence visible under the confocal microscopy<br><br> | ||
+ | - participants: Darius Rauch, Dimitri Schumacher, Elena Rätsch, Juliana Rojas Pión, Paul Herrmann" | ||
+ | data-date="July 2019" | ||
+ | data-category="chlamy"> | ||
+ | July | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="- Started with the construction of our own bioreactor. Conceptualization of the functionalities and necessary features had before been experimentally determined during our multicultivation tests<br>" | ||
+ | data-date="August 2019" | ||
+ | data-category="chlamy"> | ||
+ | August | ||
+ | </div> | ||
+ | <div class="timeline-month" | ||
+ | data-text="- Wiring and programming the sensors and LEDs on the DIY-bioreactor<br> | ||
+ | - laser-cutting of the shell and cultivation compartment of our bioreactor" | ||
+ | data-date="September 2019" | ||
+ | data-category="chlamy"> | ||
+ | September | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="infobox"> | ||
+ | <div> | ||
+ | <h3 class="infobox-date"></h3> | ||
+ | <img src="" alt="" class="infobox-image no-image" /> | ||
+ | </div> | ||
+ | <p class="infobox-text"></p> | ||
+ | </div> | ||
+ | </section> | ||
+ | </div> | ||
+ | </main> | ||
+ | |||
+ | <div class="footer-container"> | ||
+ | <footer class="width-limit"> | ||
+ | <img src="https://static.igem.org/mediawiki/2019/e/ee/T--Humboldt_Berlin--logo_black.png" class="footer-logo" alt="Chlamylicious Logo Black" /> | ||
+ | <div class="footer-right"> | ||
+ | <ul> | ||
+ | <li> | ||
+ | <a href="/Team:Humboldt_Berlin"> | ||
+ | Home | ||
+ | </a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="/Team:Humboldt_Berlin/Team"> | ||
+ | Team | ||
+ | </a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="/Team:Humboldt_Berlin/Description"> | ||
+ | Project | ||
+ | </a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | <ul> | ||
+ | <li> | ||
+ | <a href="/Team:Humboldt_Berlin/Parts"> | ||
+ | Parts | ||
+ | </a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="/Team:Humboldt_Berlin/Safety"> | ||
+ | Safety | ||
+ | </a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="/Team:Humboldt_Berlin/Human_Practices"> | ||
+ | Human Practices | ||
+ | </a> | ||
+ | </li> | ||
+ | <li> | ||
+ | <a href="https://igem.org/2019_Judging_Form?team=Humboldt_Berlin"> | ||
+ | For Judges | ||
+ | </a> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </footer> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <script> | ||
+ | var elementOffset = 160; | ||
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+ | var infobox = document.querySelector('.infobox'); | ||
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+ | |||
+ | infoboxText.innerHTML = firstElement.getAttribute('data-text'); | ||
+ | infoboxDate.innerHTML = firstElement.getAttribute('data-date'); | ||
+ | |||
+ | setFilter('synthesis'); | ||
+ | |||
+ | [].forEach.call(categoryFilter, function(el) { | ||
+ | el.addEventListener('click', function() { | ||
+ | setFilter(el.getAttribute('data-category')); | ||
+ | }); | ||
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+ | |||
+ | window.addEventListener("scroll", function () { | ||
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+ | setFixedElements(); | ||
}); | }); | ||
+ | |||
+ | function setFixedElements() { | ||
+ | var timelineOffset = document.querySelector('.timeline').getBoundingClientRect().top; | ||
− | + | if (timelineOffset < 150) { | |
− | + | infobox.classList.add('fixed') | |
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− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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} | } | ||
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− | + | container.classList.remove('bioreaktor-color'); | |
− | + | container.classList.remove('chlamy-color'); | |
− | + | container.classList.remove('human-practice-color'); | |
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+ | container.classList.add(filter + '-color'); | ||
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+ | infoboxText.innerHTML = lastActive.getAttribute('data-text') | ||
+ | infoboxDate.innerHTML = lastActive.getAttribute('data-date'); | ||
+ | |||
+ | const imageUrl = lastActive.getAttribute('data-image'); | ||
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+ | infobox.classList.remove('has-image'); | ||
+ | infoboxImage.src = ""; | ||
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</html> | </html> |
Latest revision as of 21:47, 21 October 2019
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