Difference between revisions of "Team:Toulouse-INSA-UPS/Notebook"

 
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<p> Here, you will find information about the temporal aspects of our project.</p>
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<p> Here, you will find information about the timeline of our project.</p>
 
 
 
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<ul class="nav nav-pills d-flex flex-row flex-wrap flex-lg-nowrap justify-content-around justify-content-lg-start" role="tablist">
 
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<a class="nav-link btn btn-secondary rounded-lg-top corner-lg-bottom mr-1 mb-1 mb-lg-0 active" data-toggle="pill" href="#NOTEBOOK_Part1" role="tab">LabBook</a>
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<a class="nav-link btn btn-secondary rounded-lg-top corner-lg-bottom mr-1 mb-1 mb-lg-0 active" data-toggle="pill" href="#NOTEBOOK_Part1" role="tab" style="color:white !important;" >LabBook</a>
 
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<a class="nav-link btn btn-secondary rounded-lg-top corner-lg-bottom mr-1 mb-1 mb-lg-0" data-toggle="pill" href="#NOTEBOOK_Part2" role="tab">Extra-LabBook</a>
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<a class="nav-link btn btn-secondary rounded-lg-top corner-lg-bottom mr-1 mb-1 mb-lg-0" data-toggle="pill" href="#NOTEBOOK_Part2" role="tab" style="color:white !important;">Extra-LabBook</a>
 
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part9" role="tab">Sep.</a>
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part9" role="tab" style="color:white !important;" >Sep.</a>
 
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part8" role="tab">Aug.</a>
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part8" role="tab" style="color:white !important;">Aug.</a>
 
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part7" role="tab">Jul.</a>
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part7" role="tab" style="color:white !important;">Jul.</a>
 
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part6" role="tab">Jun.</a>
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#LabBook_Calendar_Part6" role="tab" style="color:white !important;">Jun.</a>
 
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0 active" data-toggle="pill" href="#LabBook_Calendar_Part1" role="tab">LabBook</a>
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<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0 active" data-toggle="pill" href="#LabBook_Calendar_Part1" role="tab" style="color:white !important;">LabBook</a>
 
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<h2 class="heavy">LABBOOK</h2>
 
<h2 class="heavy">LABBOOK</h2>
 
<hr/>
 
<hr/>
−
<p>To achieve our lab work, we splited into different teams, each working on different aspects of the experimental procedure. Three teams were then formed :</p>
+
<p>To achieve our lab work, we split up into different teams, each working on different aspects of the experimental procedure. Three teams were then formed :</p>
 
<ul>
 
<ul>
 
<li><strong>Team coli</strong>: this team was managed by <strong>Younes</strong> and included <strong>Marion</strong>. Their goal was to achieve the production and characterization of Cerberus and Sirius in <em>Escherichia coli</em></li>
 
<li><strong>Team coli</strong>: this team was managed by <strong>Younes</strong> and included <strong>Marion</strong>. Their goal was to achieve the production and characterization of Cerberus and Sirius in <em>Escherichia coli</em></li>
 
<li><strong>Team pastoris</strong>: this team was ruled by <strong>Angeline</strong> and included <strong>Ga&euml;lle</strong>. Their mission was to reproduce the experiments planned on <em>Escherichia coli</em> in <em>Pichia pastoris</em> </li>
 
<li><strong>Team pastoris</strong>: this team was ruled by <strong>Angeline</strong> and included <strong>Ga&euml;lle</strong>. Their mission was to reproduce the experiments planned on <em>Escherichia coli</em> in <em>Pichia pastoris</em> </li>
 
<li><strong>Team biotin</strong>: this team was ruled by <strong>Amandine</strong> and included <strong>Jean</strong> and <strong>Julien</strong>. Their objective was to construct and to test our <em>in vivo</em> biotinylation system.</li>
 
<li><strong>Team biotin</strong>: this team was ruled by <strong>Amandine</strong> and included <strong>Jean</strong> and <strong>Julien</strong>. Their objective was to construct and to test our <em>in vivo</em> biotinylation system.</li>
−
<li><strong>Team callulose</strong>: this team was managed by <strong>Cal</strong> (so the coined Callulose name) and got help from <strong>Younes</strong>. Its mission was to launch and optimize our bacterial cellulose production. </li>
+
<li><strong>Team callulose</strong>: this team was managed by <strong>Callum</strong> (and so was dubbed Callulose) and got help from <strong>Younes</strong>. Its mission was to launch and optimize our bacterial cellulose production. </li>
 
</ul>
 
</ul>
 
</div>
 
</div>
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             <h4 class="heavy">18/06 - 22/06:</h4>
 
             <h4 class="heavy">18/06 - 22/06:</h4>
 
<p>
 
<p>
−
For the first week in the lab, we recovered and amplify our plasmids (pPICz, pETDuet, pET28, pGAP) but also our bacterial strains <i>E. coli</i> BL21 DE3 and Tuner.
+
For the first week in the lab, we received and amplified our plasmids (pPICz, pETDuet, pET28, pGAP) but also our bacterial strains <i>E. coli</i> BL21 DE3 and Tuner.
 
</p>
 
</p>
 
 
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             <h4 class="heavy">25/06 - 29/06:</h4>
 
             <h4 class="heavy">25/06 - 29/06:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team coli</strong>: we started with a PCR on the the gblocks Cerberus-pastoris. In order to clone this gblocks, we prepared the pGAP plasmid by amplifying it in <i>E. coli</i> Stellard cells before a Midiprep. Another aspect was to check the pREAV which will allow the incorporation of a unnatural amino acid in our Cerberus.</li>
+
<li><strong>Team coli</strong>: we started with a PCR on the the gBlock Cerberus-pastoris. In order to clone this gBlock, we prepared the pGAP plasmid by amplifying it in <i>E. coli</i> Stellar cells before a Midiprep. Another aspect was to check the pREAV which will allow the incorporation of a unnatural amino acid in our Cerberus.</li>
−
<li><strong>Team pastoris</strong>: pPIC, pGAP and pETDuet were good but pPIC did not correspond to our expectations so we had to find another solution.</li>
+
<li><strong>Team pastoris</strong>: pPIC, pGAP and pETDuet were good but pPIC did not match our expectations so we had to find another solution.</li>
 +
 
 +
<li><strong>Team callulose</strong>: the first productions of cellulose started in culture tubes.</li>
 
</ul>
 
</ul>
 
         </div>
 
         </div>
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             <h4 class="heavy">02/07 - 06/07:</h4>
 
             <h4 class="heavy">02/07 - 06/07:</h4>
 
<ul>
 
<ul>
−
<li><strong>Team biotin</strong>: this week, we have done BirA PCR and we had digested the pPIC after his amplification in <i>E. coli</i> Stellard cells. This allowed us to do our first cloning with BirA in the pETDuet. We have checked the clones by digestion and they were OK. It was necessary to sequence this clones before doing the next cloning in  pETDuet. We found another pPIC and we checked it.   
+
<li><strong>Team biotin</strong>: this week, we performed a PCR on BirA and we digested the pPIC after its amplification in <i>E. coli</i> Stellar cells. This allowed us to do our first cloning with BirA in the pETDuet. We checked the clones by digestion and they were OK. It was necessary to sequence these clones before doing the next cloning in  pETDuet. We obtained a different pPIC from a previous year's supplies, and we checked it.   
 
</li>
 
</li>
−
<li><strong>Team coli</strong>: this week, it was the first In-fusion cloning with our Cerberus in the pET28. We used two differents construction for Cerberus, one with a monomeric streptavidin and another with a  tetrameric streptavidin. Then, we checked the construct by digestion before sending it for sequencing. In the same time we checked our plasmid pEVOL-azF which allow the unnatural amino acid incorporation in <i>E. coli</i>.
+
<li><strong>Team coli</strong>: this week, it was the first In-fusion cloning with our Cerberus in the pET28. We used two differents construction for Cerberus, one with a monomeric streptavidin and another with a  tetrameric streptavidin. Then, we checked the constructs by digestion before sending them for sequencing. In the same time we checked our plasmid pEVOL-AzF which allows the unnatural amino acid incorporation in <i>E. coli</i>.
 
</li>
 
</li>
−
<li><strong>Team pastoris</strong>: this week, we did the In-fusion cloning of Cerberus-strepta in pGAP with the material prepared the week before. So we had to check some clones by digestion before sequencing. Fortunately, they were good. It was also this week that we received the <i>Pichia pastoris</i> GS200 strains cell.
+
<li><strong>Team pastoris</strong>: this week, we clone Cerberus-streptavidin though In-fusion into the pGAP with the material prepared the week before. So we had to check some clones by digestion before sequencing. Fortunately, they were good. It was also this week that we received the <i>Pichia pastoris</i> GS200 strains cell.
 
</li>
 
</li>
 
</ul>
 
</ul>
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<ul>
 
<ul>
−
<li><strong>Team biotin:</strong> on monday we sent our pETDuet BirA clones obtained last week to sequencing and we were happy that our clones were validated. In the same time, we carried on the PCR with the parts that will serve us to functionalise our Cerberus after biotinylation by BirA. </li>
+
<li><strong>Team biotin:</strong> on monday we sent our pETDuet BirA clones obtained last week to sequencing and we were happy that our clones were validated. In the same time, we carried on the PCRs with the parts that will be used to functionalize our Cerberus after biotinylation by BirA. </li>
−
<li><strong>Team coli:</strong> This week, we received the sequencing results for Cerberus and it was again a successfully cloning for the Toulouse iGEM team. We cloned Sirius (CBM3-RFP) in the pET28 with the In-fusion kit. To do this, we performed PCR on the gBlocks then we used the In-fusion kit. After verification, Sirius was OK.
+
<li><strong>Team coli:</strong> This week, we received the sequencing results for Cerberus and it was again a success for the Toulouse iGEM team. We cloned Sirius (CBM3-RFP) in the pET28 with the In-fusion kit. For this, we performed PCR on the gBlocks then we used the In-fusion kit. After verification, Sirius was OK.
 
</li>
 
</li>
−
<li><strong>Team pastoris:</strong> we checked the pREAV and found out the plasmid map didn’t correspond with our digestion results. We needed to find another pREAV... </li>
+
<li><strong>Team pastoris:</strong> we checked the pREAV and found out the plasmid map didn’t match with our digestion results. We needed to find a different pREAV... </li>
 +
 
 +
<li><strong>Team callulose:</strong> we decided to focus the production with <i>G. hansenii</i> because it produces more cellulose than <i>K. rhaeticus</i>. We also started production in 2L Erlenmeyers. </li>
 
</ul>
 
</ul>
 
 
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             <h4 class="heavy">16/07 - 20/07:</h4>
 
             <h4 class="heavy">16/07 - 20/07:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team biotin:</strong> This week, we focused on the pastoris cloning. In one hand, we prepared the pGAP vector (amplification, purification and digestion), mRFP1, BFP and Sygonadine parts and we did the three cloning. We tried to amplify and purify pPIC but unfortunately it was necessary to try again because the Midiprep didn’t succeed.</li>
+
<li><strong>Team biotin:</strong> This week, we focused on the pastoris cloning. We prepared the pGAP vector (amplification, purification and digestion), mRFP1, BFP and scygonadin parts and we cloned all three. We tried to amplify and purify pPIC but unfortunately it was necessary to try again because the Midiprep didn’t succeed.</li>
−
<li><strong>Team coli:</strong> this week, we made or own competent cells using BL21 and tuner strains</li>
+
<li><strong>Team coli:</strong> this week, we made or own competent cells using BL21 DE3 and tuner strains</li>
 
<li><strong>Team pastoris:</strong> this week, we did In-fusion cloning in the pGAP but this time with the cerberus mSA2. After transformation, culture and miniprep, we checked clones by digestion. The cloning was a success.</li>
 
<li><strong>Team pastoris:</strong> this week, we did In-fusion cloning in the pGAP but this time with the cerberus mSA2. After transformation, culture and miniprep, we checked clones by digestion. The cloning was a success.</li>
 
</ul>
 
</ul>
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             <h4 class="heavy">23/07 - 27/07:</h4>
 
             <h4 class="heavy">23/07 - 27/07:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team biotin:</strong> this week, before another midiprep, we checked the pPIC but we still had a mapping problem. After many verification, it was clearly not good but we found another source for this plasmid and it seemed OK. We amplified it by midiprep.</li>
+
<li><strong>Team biotin:</strong> this week, before another midiprep, we checked the pPIC but we still had a mapping problem. After many verifications, it was clearly not correct but we found another source for this plasmid and it seemed OK. We amplified it by midiprep.</li>
 
<li><strong>Team coli:</strong> this week, we did the first production assay for Sirius and we were very happy because we obtained a red culture medium.</li>
 
<li><strong>Team coli:</strong> this week, we did the first production assay for Sirius and we were very happy because we obtained a red culture medium.</li>
 
<li><strong>Team pastoris:</strong> this week, the pichia team helped the other teams because it was ahead of the planning and had to wait for yeast growth.</li>
 
<li><strong>Team pastoris:</strong> this week, the pichia team helped the other teams because it was ahead of the planning and had to wait for yeast growth.</li>
 +
 
</ul>
 
</ul>
 
         </div>
 
         </div>
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             <h4 class="heavy">30/07 - 03/08:</h4>
 
             <h4 class="heavy">30/07 - 03/08:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team biotin:</strong> this week, we worked on pETDuet-BirA to clone our functionalising proteins (mRFP1 and BFP). We amplified the pETDuet-BirA and after Midiprep and digestion, we cloned mRFP1 and BFP with the In-fusion kit. After digestion, we sent clones to sequencing and started production assay in <i>E. coli</i> BL21.</li>
+
<li><strong>Team biotin:</strong> this week, we worked on pETDuet-BirA to clone our functionalizing proteins (mRFP1 and BFP). We amplified the pETDuet-BirA and after Midiprep and digestion, we cloned mRFP1 and BFP with the In-fusion kit. After digestion, we sent clones to sequencing and started production assay in <i>E. coli</i> BL21 DE3.</li>
−
<li><strong>Team coli:</strong> we did the first Sirius purification after production in <i>E. coli</i> BL21. To do this, we used an IMAC column and dialysis. In order to proved the CBM3 fixation to cellulose, we prepared the regenerated amorphous cellulose.</li>
+
<li><strong>Team coli:</strong> we did the first Sirius purification after production in <i>E. coli</i> BL21 DE3. To do this, we used an IMAC column and dialysis. In order to prove the CBM3 fixation to cellulose, we prepared the regenerated amorphous cellulose.</li>
 +
 
 +
<li><strong>Team callulose:</strong> We intensified the cellulose production to be able to give 5g of dried cellulose to the Bordeaux iGEM team at the end of the month.</li>
 
 
 
</ul>
 
</ul>
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             <h4 class="heavy">06/08 - 10/08:</h4>
 
             <h4 class="heavy">06/08 - 10/08:</h4>
 
<ul>
 
<ul>
−
<li><strong>Team biotin</strong>: this week, we worked on the production assay for biotinylated proteins in <i>E. coli</i>. Furthermore, we cloned BirA in the pPIC plasmid and we checked it by digestion and sequencing. After that, we cloned RFP, Sygonadine and BFP in this plasmid to produce the proteins in <i>Pichia pastoris</i>.
+
<li><strong>Team biotin</strong>: this week, we worked on the production assay for biotinylated proteins in <i>E. coli</i>. Furthermore, we cloned BirA in the pPIC plasmid and we checked it by digestion and sequencing. After that, we cloned RFP, scygonadin and BFP in this plasmid to produce the proteins in <i>Pichia pastoris</i>.
 
</li>
 
</li>
 
<li><strong>Team coli:</strong> in order to show the efficiency of the second Cerberus head, we needed a Cerberus which has just its CBM3a head and the streptavidin head. We did a directed mutagenesis on the stop codon to create Orthos.</li>
 
<li><strong>Team coli:</strong> in order to show the efficiency of the second Cerberus head, we needed a Cerberus which has just its CBM3a head and the streptavidin head. We did a directed mutagenesis on the stop codon to create Orthos.</li>
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             <h4 class="heavy">20/08 - 24/08:</h4>
 
             <h4 class="heavy">20/08 - 24/08:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team coli:</strong> this week was a particularly busy week. We did the Orthos production and purification but also the same for Sirius and mRFP1. This allowed us to perform a cellulose pull down assay with Sirius and mRFP1 as a negative control. In addition, we did the production of Cerberus. To do this, we did co-transformation of BL21 cells with pET28 Cerberus and pEVOL AzF. The cell culture medium was complemented with with AzF unnatural amino acid to allow its incorporation. Cerberus was purified by a cellulose pool down assay.</li>
+
<li><strong>Team coli:</strong> this week was a particularly busy week. We produced and purified Orthos as well as for Sirius and mRFP1. This allowed us to perform a cellulose pull down assay with Sirius and mRFP1 as a negative control. In addition, we produced Cerberus. To do this, we co-transformed BL21 DE3 cells with pET28 Cerberus and pEVOL AzF. The cell culture medium was complemented with the AzF unnatural amino acid to allow its incorporation. Cerberus was purified by a cellulose pull-down assay.</li>
−
<li><strong>Team pSB1C3:</strong> we created this new team to start the pSB1C3 clonings. PCR was performed on our gblocks. Unfortunately, we didn’t succeed to amplify all the gblocks.</li>
+
<li><strong>Team pSB1C3:</strong> we created this new team to start the pSB1C3 clonings. PCRs were performed on our gBlocks. Unfortunately, we didn’t manage to amplify all the gBlocks.</li>
 
</ul>
 
</ul>
 
         </div>
 
         </div>
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             <h4 class="heavy">27/08 - 31/08:</h4>
 
             <h4 class="heavy">27/08 - 31/08:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team coli:</strong> in order to demonstrate the efficiency of the third Cerberus head, we did a click assay with fluorescein DBCO.</li>
+
<li><strong>Team coli:</strong> in order to demonstrate the efficiency of the third Cerberus head, we did a click assay with DBCO-fluorescein.</li>
 
</ul>
 
</ul>
 
         </div>
 
         </div>
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             <h4 class="heavy">03/09 - 07/09:</h4>
 
             <h4 class="heavy">03/09 - 07/09:</h4>
 
<ul>
 
<ul>
−
<li><strong>Team coli:</strong> in order to demonstrate the efficiency of the AzF head, we tried to click paramagnetic beads on it. The aim was to see the displacement of the cellulose when in a presence of a magnet</li>
+
<li><strong>Team coli:</strong> in order to demonstrate the efficiency of the AzF head, we tried to click paramagnetic beads on it. The aim was to see the movement of cellulose in a presence of a magnet</li>
−
<li><strong>Team pSB1C3:</strong> this week, we tried again the pSB1C3 clonings. We used another pSB1C3 and tried a classic cloning with restriction enzymes instead of In-fusion. At the end of the week, we sent our pSB1C3 to sequencing. </li>
+
<li><strong>Team pSB1C3:</strong> this week, we tried the pSB1C3 clonings again. We used another pSB1C3 and tried a classic cloning with restriction enzymes instead of In-fusion. At the end of the week, we sent our pSB1C3 for sequencing. </li>
 
</ul>
 
</ul>
  
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             <h4 class="heavy">10/09 - 14/09: </h4>
 
             <h4 class="heavy">10/09 - 14/09: </h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team biotin:</strong> we successfully made a pull down assay with Orthos to measure the BFP fluorescence with and without Orthos on cellulose.</li>
+
<li><strong>Team biotin:</strong> we successfully pulled Orthos down to measure the BFP fluorescence with and without Orthos on cellulose.</li>
 
</ul>
 
</ul>
 
 
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             <h4 class="heavy">17/09 - 21/09:</h4>
 
             <h4 class="heavy">17/09 - 21/09:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team biotin:</strong> this was the final assay and we proved the fixation of the biotinylated BFP on cerberus.</li>
+
<li><strong>Team biotin:</strong> this was the final assay and we proved the fixation of the biotinylated BFP on Cerberus.</li>
 +
 
 
 
 
</ul>
 
</ul>
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             <h4 class="heavy">24/09 - 28/09:</h4>
 
             <h4 class="heavy">24/09 - 28/09:</h4>
 
             <ul>
 
             <ul>
−
<li><strong>Team coli</strong>: for our last lab week, we did two manipulations, the first to show that we can fix BFP-DBCO to the head streptavidin and the second to test the specificity of CBM3a to bacterial cellulose. And we successfully validated these two experiments!</li>
+
<li><strong>Team coli & callulose</strong>: for our last lab week, we managed to produce a pink bacterial cellulose by using our purified Sirius protein.</li>
 
</ul>
 
</ul>
 
 
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<ul class="nav nav-pills d-flex flex-row-reverse flex-wrap flex-lg-nowrap justify-content-around justify-content-lg-start mb-2" role="tablist">
 
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−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part10" role="tab">Oct.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part10" role="tab" style="color:white !important;">Oct.</a>
 
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</li>
 
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<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part9" role="tab">Sep.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part9" role="tab" style="color:white !important;">Sep.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part8" role="tab">Aug.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part8" role="tab" style="color:white !important;">Aug.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part7" role="tab">Jul.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part7" role="tab" style="color:white !important;">Jul.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part6" role="tab">Jun.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part6" role="tab" style="color:white !important;">Jun.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part5" role="tab">May</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part5" role="tab" style="color:white !important;">May</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part4" role="tab">Apr.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part4" role="tab" style="color:white !important;">Apr.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part3" role="tab">Mar.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part3" role="tab" style="color:white !important;">Mar.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part2" role="tab">Feb.</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0" data-toggle="pill" href="#OtherBook_Calendar_Part2" role="tab" style="color:white !important;">Feb.</a>
 
</li>
 
</li>
 
<li class="nav-item">
 
<li class="nav-item">
−
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0 active" data-toggle="pill" href="#OtherBook_Calendar_Part1" role="tab">Extra-LabBook</a>
+
<a class="nav-link btn btn-secondary rounded-lg-bottom corner-lg-top ml-1 mt-1 mt-lg-0 active" data-toggle="pill" href="#OtherBook_Calendar_Part1" role="tab" style="color:white !important;">Extra-LabBook</a>
 
</li>
 
</li>
 
</ul>
 
</ul>
 
 
−
<!--INFORMATIVE CONTENT--> <p>Here, you can find about some of our activities outside of the lab. More details are provided in the <a href="https://2018.igem.org/Team:Toulouse-INSA-UPS/Human_Practices">HP LogBOOK</a> of the Integrated Human Practices section.</p>
+
<!--INFORMATIVE CONTENT-->
 
<div class="tab-content">
 
<div class="tab-content">
 
<div class="tab-pane fade show active" id="OtherBook_Calendar_Part1" role="tabpanel">
 
<div class="tab-pane fade show active" id="OtherBook_Calendar_Part1" role="tabpanel">
−
<h2 class="heavy">Extra-LabBOOK</h2>
+
<h2 class="heavy">Extra-LabBook</h2>
 
<hr/>
 
<hr/>
−
+
<p>Here, you can find about some of our activities outside of the lab. More details are provided in the <a href="https://2018.igem.org/Team:Toulouse-INSA-UPS/Human_Practices">HP LogBook</a> of the Integrated Human Practices section.</p>
 
</div>
 
</div>
 
<!--End : PART 2.1-->
 
<!--End : PART 2.1-->
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             <h4 class="heavy">February 1st, 2018: Kick-off meeting </h4>
 
             <h4 class="heavy">February 1st, 2018: Kick-off meeting </h4>
 
<p>
 
<p>
−
Gathering the team members, distribute the tasks and of course get to know each others…
+
First meeting of the team, distribution of the tasks and of course we got to know each other.
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/5/54/T--Toulouse-INSA-UPS--NB--Youn--Team.jpg" style="width:60%" alt=""/>
 
<img src="https://static.igem.org/mediawiki/2018/5/54/T--Toulouse-INSA-UPS--NB--Youn--Team.jpg" style="width:60%" alt=""/>
 
<p>
 
<p>
−
The iGEM’s adventure begins!  
+
The iGEM adventure began!  
 
</p>
 
</p>
  
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             <h4 class="heavy">February 8th-22nd, 2018: Brainstorming </h4>
 
             <h4 class="heavy">February 8th-22nd, 2018: Brainstorming </h4>
 
             <p>
 
             <p>
−
At the beginning, more than 50 ideas were proposed. The most original ideas were selected during our brainstorming sessions.  
+
At the beginning, more than 50 ideas were suggested. The most original ideas were selected during our brainstorming sessions.  
 
</p>
 
</p>
 
<p>
 
<p>
−
At the end of the month, still 11 subjects need more investigations.
+
At the end of the month, 11 subjects still needed more investigations.
 
</p>
 
</p>
 
 
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             <h4 class="heavy">February 21st, 2018: Escape game conference</h4>
 
             <h4 class="heavy">February 21st, 2018: Escape game conference</h4>
 
             <p>
 
             <p>
−
We attend a conference organized by the Catalyseur on the creation of a teaching escape game. We met two professors in high schools option “Sciences and technologies laboratory”.
+
We attended a conference organized by Le Catalyseur on the creation of a teaching escape game. We met two professors in high schools option “Sciences and Technologies of the Laboratory” (STL).
 
</p>
 
</p>
 
         </div>
 
         </div>
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">March 1st-15t, 2018: Brainstorming</h4>
+
             <h4 class="heavy">March 1st-15th, 2018: Brainstorming</h4>
 
<p>
 
<p>
−
Only 5 subjects still in course!  
+
Only 5 subjects left!  
 
</p>
 
</p>
  
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             <h4 class="heavy">March 10th, 2018: Grimoire</h4>
 
             <h4 class="heavy">March 10th, 2018: Grimoire</h4>
 
             <p>
 
             <p>
−
At this event we introduced our card game Microbioworld.
+
At this event we introduced our card game, Microbioworld.
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/a/aa/T--Toulouse-INSA-UPS--NB--Youn--grim.jpg" style="width:60%" alt=""/>
 
<img src="https://static.igem.org/mediawiki/2018/a/aa/T--Toulouse-INSA-UPS--NB--Youn--grim.jpg" style="width:60%" alt=""/>
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             <h4 class="heavy">March 20th, 2018: CRISPR/CAS meeting</h4>
 
             <h4 class="heavy">March 20th, 2018: CRISPR/CAS meeting</h4>
 
             <p>
 
             <p>
−
We meet a CRISPR/CAS expert in the LMGM lab who teaches us a lot about the new possibilities of the system.  
+
We met a CRISPR/CAS expert in the LMGM lab who taught us a lot about the new possibilities of the system.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/2/2b/T--Toulouse-INSA-UPS--NB--Youn--crispr.jpg" style="width:60%" alt=""/>
 
<img src="https://static.igem.org/mediawiki/2018/2/2b/T--Toulouse-INSA-UPS--NB--Youn--crispr.jpg" style="width:60%" alt=""/>
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             <h4 class="heavy">April 5th, 2018: Final choice of our project </h4>
 
             <h4 class="heavy">April 5th, 2018: Final choice of our project </h4>
 
<p>
 
<p>
−
After a long period of brainstorming, we have decided to work on molecules’ binding on cellulose. Now, we had to think of our project’s name.
+
After a long period of brainstorming, we decided to work on the binding of molecules on cellulose. Then, we had to think about the name of our project.
 
</p>
 
</p>
  
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">May , 2018: </h4>
+
             <h4 class="heavy">May 10th, 2018: </h4>
 
<p>
 
<p>
 
We worked on the design of the project with the help of our supervisors.  
 
We worked on the design of the project with the help of our supervisors.  
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">May 31th, 2018: </h4>
+
             <h4 class="heavy">May 31st, 2018: </h4>
 
             <p>
 
             <p>
−
We met Philippe Serp and his team to manipulate our graphene on his lab.  
+
We met Philippe Serp and his team to talk about experimenting in his lab. We needed them to activate our graphene before using it to functionalise cellulose.
 
</p>
 
</p>
 
 
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             <h4 class="heavy">June 18th, 2018: First day in the lab</h4>
 
             <h4 class="heavy">June 18th, 2018: First day in the lab</h4>
 
<p>
 
<p>
−
We were all very impatient, and finally it has come! It’s the first lab day.  
+
We were all very impatient, and finally it has come! It’s the first day of labwork.  
 
</p>
 
</p>
  
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">June 19th, 2018: Fondation INSA Toulouse meeting</h4>
+
             <h4 class="heavy">June 19th, 2018: Foundation INSA Toulouse meeting</h4>
 
             <p>
 
             <p>
−
Looking for funds, we presented our project to the INSA Toulouse fondation.
+
Looking for funds, we presented our project to the INSA Toulouse foundation.
 
</p>
 
</p>
 
 
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">June 23rd-24th, 2018: Collaboration with iGEM Montpellier’s team</h4>
+
             <h4 class="heavy">June 23rd-24th, 2018: Collaboration with the Montpellier iGEM team</h4>
 
             <p>
 
             <p>
−
We starte a collaboration with the Montpellier team to help them in some areas like the wiki. To do so, we went to Montpellier.  
+
We started a collaboration with the Montpellier team to help them in some areas like the wiki. To do so, we went to Montpellier to talk with them and share our knowledge of the competition.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/6/60/T--Toulouse-INSA-UPS--NB--Youn--mtp.jpg" style="width:60%" alt=""/>
 
<img src="https://static.igem.org/mediawiki/2018/6/60/T--Toulouse-INSA-UPS--NB--Youn--mtp.jpg" style="width:60%" alt=""/>
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             <h4 class="heavy">June 25th, 2018: TWB meeting</h4>
 
             <h4 class="heavy">June 25th, 2018: TWB meeting</h4>
 
             <p>
 
             <p>
−
We have met Pierre Monsan in Toulouse White Biotechnology (TWB), one of our main funders.  
+
We met Pierre Monsan, one of the main founders of one of our sponsors, Toulouse White Biotechnology (TWB).  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/a/a6/T--Toulouse-INSA-UPS--NB--Youn--twb.jpg" style="width:60%" alt=""/>
 
<img src="https://static.igem.org/mediawiki/2018/a/a6/T--Toulouse-INSA-UPS--NB--Youn--twb.jpg" style="width:60%" alt=""/>
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             <h4 class="heavy">June 26th, 2018: NUS meeting</h4>
 
             <h4 class="heavy">June 26th, 2018: NUS meeting</h4>
 
             <p>
 
             <p>
−
We organized a skype meeting with the NUS iGEM team to discuss about a possible collaboration.  
+
We organized a Skype meeting with the NUS iGEM team to discuss about a possible collaboration.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/9/95/T--Toulouse-INSA-UPS--NB--Youn--nusskype.jpg" alt="" style="width:60%"/>
 
<img src="https://static.igem.org/mediawiki/2018/9/95/T--Toulouse-INSA-UPS--NB--Youn--nusskype.jpg" alt="" style="width:60%"/>
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">June 27th, 2018: Meet with STL teachers</h4>
+
             <h4 class="heavy">June 27th, 2018: Meeting the STL teachers</h4>
 
             <p>
 
             <p>
−
As part of our discussions with the ministry of education, high school teachers visited the lab where we worked during summer.  
+
As part of our discussions with the french ministry of education, high school teachers visited the lab where we worked during summer.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/1/1f/T--Toulouse-INSA-UPS--NB--Youn--stlvisit.jpg" alt="" style="width:60%"/>
 
<img src="https://static.igem.org/mediawiki/2018/1/1f/T--Toulouse-INSA-UPS--NB--Youn--stlvisit.jpg" alt="" style="width:60%"/>
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             <h4 class="heavy">June 27th, 2018</h4>
 
             <h4 class="heavy">June 27th, 2018</h4>
 
             <p>
 
             <p>
−
Entrep: We met Guillaume Boissonat, who co-funded the startup pili.bio. He shared with us his own experience as a co-funder, and he gave to us some advice about the possible business model for Cerberus Startup.  
+
For our entrepreneurship approach, we met Guillaume Boissonat who co-funded the startup pili.bio. He shared with us his own experience as a co-funder, and he gave us some advice about the possible business model for a Cerberus startup.  
 
</p>
 
</p>
 
 
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             <h4 class="heavy">June 29th, 2018: Primary school intervention</h4>
 
             <h4 class="heavy">June 29th, 2018: Primary school intervention</h4>
 
             <p>
 
             <p>
−
We intervened in a primary school in order to present Microbiolworld, the research profession, and more generally introduce the world of microbes to children.  
+
We gave a class in a primary school in order to present Microbiolworld, the research profession, and more generally to introduce the world of microbes to children.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/1/19/T--Toulouse-INSA-UPS--NB--Youn--school.jpg" alt="" style="width:60%"/>
 
<img src="https://static.igem.org/mediawiki/2018/1/19/T--Toulouse-INSA-UPS--NB--Youn--school.jpg" alt="" style="width:60%"/>
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             <h4 class="heavy">July 5th, 2018: CALMIP’s meet and visit</h4>
 
             <h4 class="heavy">July 5th, 2018: CALMIP’s meet and visit</h4>
 
<p>
 
<p>
−
We visited CALMIP a high-throughput calcul center which allowed us to make complex calculations for our modelling.  
+
We visited CALMIP, a high-throughput calculations center which allowed us to execute complex algorithms for our modelling.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/3/3a/T--Toulouse-INSA-UPS--NB--Youn--calmip.jpg" alt="" style="width:50%"/>
 
<img src="https://static.igem.org/mediawiki/2018/3/3a/T--Toulouse-INSA-UPS--NB--Youn--calmip.jpg" alt="" style="width:50%"/>
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             <h4 class="heavy">July 6th, 2018: Official announcement of our project</h4>
 
             <h4 class="heavy">July 6th, 2018: Official announcement of our project</h4>
 
             <p>
 
             <p>
−
We officially announced the purpose of our project on social media!  
+
We officially announced the purpose of our project on social medias!  
 
</p>
 
</p>
 
 
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         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">July 7th, 2018: 4th Annual Parisian Meet-up</h4>
+
             <h4 class="heavy">July 7th, 2018: 4th Annual parisian meet-up</h4>
 
             <p>
 
             <p>
−
We took part in the annual Parisian Meet-up organized by Pasteur iGEM team.  
+
We took part in the annual parisian meet-up organized by the Pasteur iGEM team.  
 
</p>
 
</p>
 
<img style="margin-right:25px; margin-bottom:2px;" src="https://static.igem.org/mediawiki/2018/4/4a/T--Toulouse-INSA-UPS--Collaborations--GB--MeetupParis.jpg" alt="Meet up Paris" width="52%" height="90%">
 
<img style="margin-right:25px; margin-bottom:2px;" src="https://static.igem.org/mediawiki/2018/4/4a/T--Toulouse-INSA-UPS--Collaborations--GB--MeetupParis.jpg" alt="Meet up Paris" width="52%" height="90%">
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             <h4 class="heavy">July 7th-12th, 2018: EuroScience Open Forum (ESOF)</h4>
 
             <h4 class="heavy">July 7th-12th, 2018: EuroScience Open Forum (ESOF)</h4>
 
             <p>
 
             <p>
−
We participated to the EuroScience Open Forum in Toulouse to introduce the iGEM competition and synthetic biology to the public.   
+
We participated in the EuroScience Open Forum in Toulouse to introduce the iGEM competition and synthetic biology to the public.   
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/5/52/T--Toulouse-INSA-UPS--NB--Youn--esof.jpg" style="width:60%" alt=""/>
 
<img src="https://static.igem.org/mediawiki/2018/5/52/T--Toulouse-INSA-UPS--NB--Youn--esof.jpg" style="width:60%" alt=""/>
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         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">July 19th, 2018: First publications in media</h4>
+
             <h4 class="heavy">July 19th, 2018: First publications in french newspapers</h4>
 
             <p>
 
             <p>
−
Toulouse media spoke about us and Cerberus, we were very happy of this recognition!   
+
Newspapers from Toulouse interviewed us about our Cerberus project, we were very happy and grateful for this recognition!   
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/c/c7/T--Toulouse-INSA-UPS--NB--Youn--journals.jpg" alt="" style="width:60%"/>
 
<img src="https://static.igem.org/mediawiki/2018/c/c7/T--Toulouse-INSA-UPS--NB--Youn--journals.jpg" alt="" style="width:60%"/>
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">July 20th-22nd, 2018: European Meet-up in Munich</h4>
+
             <h4 class="heavy">July 20th-22nd, 2018: European meet-up in Munich</h4>
 
             <p>
 
             <p>
 
Two members of the team travelled to Munich to represent our team at the European meet-up.
 
Two members of the team travelled to Munich to represent our team at the European meet-up.
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             <h4 class="heavy">August 1st, 2018: Laval skype</h4>
 
             <h4 class="heavy">August 1st, 2018: Laval skype</h4>
 
<p>
 
<p>
−
We contacted the laval iGEM team in order to create an innovative human practice collaboration.  
+
We contacted the Laval iGEM team in order to create an innovative human practices collaboration.  
 
</p>
 
</p>
  
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<!--Begin : BLOCK CONTENT-->
 
<!--Begin : BLOCK CONTENT-->
 
         <div class="cd-timeline__content js-cd-content">
 
         <div class="cd-timeline__content js-cd-content">
−
             <h4 class="heavy">August 17th, 2018: Shooting photo for the wiki</h4>
+
             <h4 class="heavy">August 17th, 2018: Photoshoot for the wiki</h4>
 
             <p>
 
             <p>
−
We dressed up to take the pictures used in our wiki.  
+
We dressed up to take the photos for our wiki.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/1/1c/T--Toulouse-INSA-UPS--NB--Youn--team.jpg" style="width:60%" alt="">
 
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             <h4 class="heavy">August 25th-26th, 2018: Collaboration with Bordeaux’s team</h4>
+
             <h4 class="heavy">August 25th-26th, 2018: Collaboration with Bordeaux iGEM team</h4>
 
             <p>
 
             <p>
−
We went to Bordeaux to bring them our bacterial cellulose. Then, they will use it as another carbon source for there Far Waste project.
+
We went to Bordeaux to bring them our bacterial cellulose, so they could use it as another carbon source for their Far Waste project.
 
</p>
 
</p>
 
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             <h4 class="heavy">August 29th, 2018: First working group with Le Catalyseur</h4>
 
             <h4 class="heavy">August 29th, 2018: First working group with Le Catalyseur</h4>
 
             <p>
 
             <p>
−
<i>Le Catalyseur</i> is a business incubator, they helped us to write our business plan. This first working group allowed us to structure our company and defined our objectives.  
+
<i>Le Catalyseur</i> is a business incubator, they helped us to write our business plan. This first session allowed us to structure our company and define our objectives.  
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/5/59/T--Toulouse-INSA-UPS--NB--Youn--cat.jpg" alt="" style="width:60%"/>
 
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             <h4 class="heavy">August 31st, 2018: Second workshop with <i>Le Catalyseur</i></h4>
 
             <h4 class="heavy">August 31st, 2018: Second workshop with <i>Le Catalyseur</i></h4>
 
             <p>
 
             <p>
−
During this second meeting, we identified the critical points and risk factors we need to enhance to conducted Cerberus as a business.  
+
During this second meeting, we identified the critical points and risk factors we needed to enhance to conduct Cerberus as a business.  
 
</p>
 
</p>
 
</div>
 
</div>
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             <h4 class="heavy">September 5th, 2018: </h4>
 
             <h4 class="heavy">September 5th, 2018: </h4>
 
             <p>
 
             <p>
−
<i>Le Catalyseur</i> helped us to create a SWOT matrice to define our strengths and weakness as a business.  
+
<i>Le Catalyseur</i> helped us to create a SWOT matrix to define our strengths and weaknesses as a business.  
 
</p>
 
</p>
 
 
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             <h4 class="heavy">October 3rd, 2018: </h4>
 
<p>
 
<p>
−
We have done a general repetition in an international high school near Toulouse.  
+
We presented our project in an international high school near Toulouse. This also served as a general repetition for our presentation at the giant Jamboree.
 
</p>
 
</p>
 
<img src="https://static.igem.org/mediawiki/2018/3/30/T--Toulouse-INSA-UPS--NB--Youn--ihs.jpg" alt="" style="width:60%"/>
 
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             <h4 class="heavy">October 5th, 2018: </h4>
 
             <h4 class="heavy">October 5th, 2018: </h4>
 
             <p>
 
             <p>
−
Another meeting with <i>Le Catalyseur</i> to talk about industrial property.  
+
We discussed industrial property during another meeting with <i>Le Catalyseur</i>.  
 
</p>
 
</p>
 
 
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 +
            <p>
 +
We managed to complete all our wiki pages a day earlier, with the precious help of our supervisor Brice. We had to work late into the night, but it was worth it!
 +
</p>
 +
<div class="center">
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Latest revision as of 05:36, 17 October 2018

NOTEBOOK

Here, you will find information about the timeline of our project.

LABBOOK

To achieve our lab work, we split up into different teams, each working on different aspects of the experimental procedure. Three teams were then formed :

  • Team coli: this team was managed by Younes and included Marion. Their goal was to achieve the production and characterization of Cerberus and Sirius in Escherichia coli
  • Team pastoris: this team was ruled by Angeline and included Gaëlle. Their mission was to reproduce the experiments planned on Escherichia coli in Pichia pastoris
  • Team biotin: this team was ruled by Amandine and included Jean and Julien. Their objective was to construct and to test our in vivo biotinylation system.
  • Team callulose: this team was managed by Callum (and so was dubbed Callulose) and got help from Younes. Its mission was to launch and optimize our bacterial cellulose production.

LABBOOK: JUNE

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18/06 - 22/06:

For the first week in the lab, we received and amplified our plasmids (pPICz, pETDuet, pET28, pGAP) but also our bacterial strains E. coli BL21 DE3 and Tuner.

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25/06 - 29/06:

  • Team coli: we started with a PCR on the the gBlock Cerberus-pastoris. In order to clone this gBlock, we prepared the pGAP plasmid by amplifying it in E. coli Stellar cells before a Midiprep. Another aspect was to check the pREAV which will allow the incorporation of a unnatural amino acid in our Cerberus.
  • Team pastoris: pPIC, pGAP and pETDuet were good but pPIC did not match our expectations so we had to find another solution.
  • Team callulose: the first productions of cellulose started in culture tubes.

LABBOOK: JULY

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02/07 - 06/07:

  • Team biotin: this week, we performed a PCR on BirA and we digested the pPIC after its amplification in E. coli Stellar cells. This allowed us to do our first cloning with BirA in the pETDuet. We checked the clones by digestion and they were OK. It was necessary to sequence these clones before doing the next cloning in pETDuet. We obtained a different pPIC from a previous year's supplies, and we checked it.
  • Team coli: this week, it was the first In-fusion cloning with our Cerberus in the pET28. We used two differents construction for Cerberus, one with a monomeric streptavidin and another with a tetrameric streptavidin. Then, we checked the constructs by digestion before sending them for sequencing. In the same time we checked our plasmid pEVOL-AzF which allows the unnatural amino acid incorporation in E. coli.
  • Team pastoris: this week, we clone Cerberus-streptavidin though In-fusion into the pGAP with the material prepared the week before. So we had to check some clones by digestion before sequencing. Fortunately, they were good. It was also this week that we received the Pichia pastoris GS200 strains cell.
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09/07 - 13/07:

  • Team biotin: on monday we sent our pETDuet BirA clones obtained last week to sequencing and we were happy that our clones were validated. In the same time, we carried on the PCRs with the parts that will be used to functionalize our Cerberus after biotinylation by BirA.
  • Team coli: This week, we received the sequencing results for Cerberus and it was again a success for the Toulouse iGEM team. We cloned Sirius (CBM3-RFP) in the pET28 with the In-fusion kit. For this, we performed PCR on the gBlocks then we used the In-fusion kit. After verification, Sirius was OK.
  • Team pastoris: we checked the pREAV and found out the plasmid map didn’t match with our digestion results. We needed to find a different pREAV...
  • Team callulose: we decided to focus the production with G. hansenii because it produces more cellulose than K. rhaeticus. We also started production in 2L Erlenmeyers.
icon

16/07 - 20/07:

  • Team biotin: This week, we focused on the pastoris cloning. We prepared the pGAP vector (amplification, purification and digestion), mRFP1, BFP and scygonadin parts and we cloned all three. We tried to amplify and purify pPIC but unfortunately it was necessary to try again because the Midiprep didn’t succeed.
  • Team coli: this week, we made or own competent cells using BL21 DE3 and tuner strains
  • Team pastoris: this week, we did In-fusion cloning in the pGAP but this time with the cerberus mSA2. After transformation, culture and miniprep, we checked clones by digestion. The cloning was a success.
icon

23/07 - 27/07:

  • Team biotin: this week, before another midiprep, we checked the pPIC but we still had a mapping problem. After many verifications, it was clearly not correct but we found another source for this plasmid and it seemed OK. We amplified it by midiprep.
  • Team coli: this week, we did the first production assay for Sirius and we were very happy because we obtained a red culture medium.
  • Team pastoris: this week, the pichia team helped the other teams because it was ahead of the planning and had to wait for yeast growth.
icon

30/07 - 03/08:

  • Team biotin: this week, we worked on pETDuet-BirA to clone our functionalizing proteins (mRFP1 and BFP). We amplified the pETDuet-BirA and after Midiprep and digestion, we cloned mRFP1 and BFP with the In-fusion kit. After digestion, we sent clones to sequencing and started production assay in E. coli BL21 DE3.
  • Team coli: we did the first Sirius purification after production in E. coli BL21 DE3. To do this, we used an IMAC column and dialysis. In order to prove the CBM3 fixation to cellulose, we prepared the regenerated amorphous cellulose.
  • Team callulose: We intensified the cellulose production to be able to give 5g of dried cellulose to the Bordeaux iGEM team at the end of the month.

LABBOOK: AUGUST

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06/08 - 10/08:

  • Team biotin: this week, we worked on the production assay for biotinylated proteins in E. coli. Furthermore, we cloned BirA in the pPIC plasmid and we checked it by digestion and sequencing. After that, we cloned RFP, scygonadin and BFP in this plasmid to produce the proteins in Pichia pastoris.
  • Team coli: in order to show the efficiency of the second Cerberus head, we needed a Cerberus which has just its CBM3a head and the streptavidin head. We did a directed mutagenesis on the stop codon to create Orthos.
icon

13/08 - 17/08:

  • Team biotin: we produced biotinylated proteins with E. coli, with and without biotin in the culture medium.
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20/08 - 24/08:

  • Team coli: this week was a particularly busy week. We produced and purified Orthos as well as for Sirius and mRFP1. This allowed us to perform a cellulose pull down assay with Sirius and mRFP1 as a negative control. In addition, we produced Cerberus. To do this, we co-transformed BL21 DE3 cells with pET28 Cerberus and pEVOL AzF. The cell culture medium was complemented with the AzF unnatural amino acid to allow its incorporation. Cerberus was purified by a cellulose pull-down assay.
  • Team pSB1C3: we created this new team to start the pSB1C3 clonings. PCRs were performed on our gBlocks. Unfortunately, we didn’t manage to amplify all the gBlocks.
icon

27/08 - 31/08:

  • Team coli: in order to demonstrate the efficiency of the third Cerberus head, we did a click assay with DBCO-fluorescein.

LABBOOK: SEPTEMBER

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03/09 - 07/09:

  • Team coli: in order to demonstrate the efficiency of the AzF head, we tried to click paramagnetic beads on it. The aim was to see the movement of cellulose in a presence of a magnet
  • Team pSB1C3: this week, we tried the pSB1C3 clonings again. We used another pSB1C3 and tried a classic cloning with restriction enzymes instead of In-fusion. At the end of the week, we sent our pSB1C3 for sequencing.
icon

10/09 - 14/09:

  • Team biotin: we successfully pulled Orthos down to measure the BFP fluorescence with and without Orthos on cellulose.
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17/09 - 21/09:

  • Team biotin: this was the final assay and we proved the fixation of the biotinylated BFP on Cerberus.
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24/09 - 28/09:

  • Team coli & callulose: for our last lab week, we managed to produce a pink bacterial cellulose by using our purified Sirius protein.

Extra-LabBook

Here, you can find about some of our activities outside of the lab. More details are provided in the HP LogBook of the Integrated Human Practices section.

Extra-LabBOOK: FEBRUARY

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February 1st, 2018: Kick-off meeting

First meeting of the team, distribution of the tasks and of course we got to know each other.

The iGEM adventure began!

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February 8th-22nd, 2018: Brainstorming

At the beginning, more than 50 ideas were suggested. The most original ideas were selected during our brainstorming sessions.

At the end of the month, 11 subjects still needed more investigations.

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February 21st, 2018: Escape game conference

We attended a conference organized by Le Catalyseur on the creation of a teaching escape game. We met two professors in high schools option “Sciences and Technologies of the Laboratory” (STL).

Extra-LabBOOK: MARCH

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March 1st-15th, 2018: Brainstorming

Only 5 subjects left!

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March 10th, 2018: Grimoire

At this event we introduced our card game, Microbioworld.

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March 20th, 2018: CRISPR/CAS meeting

We met a CRISPR/CAS expert in the LMGM lab who taught us a lot about the new possibilities of the system.

Extra-LabBOOK: APRIL

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April 5th, 2018: Final choice of our project

After a long period of brainstorming, we decided to work on the binding of molecules on cellulose. Then, we had to think about the name of our project.

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April 12th, 2018: Tour des Sciences

We presented the Microbioworld game during the ”Tour des sciences”.

Extra-LabBOOK: MAY

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May 10th, 2018:

We worked on the design of the project with the help of our supervisors.

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May 31st, 2018:

We met Philippe Serp and his team to talk about experimenting in his lab. We needed them to activate our graphene before using it to functionalise cellulose.

Extra-LabBOOK: JUNE

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June 18th, 2018: First day in the lab

We were all very impatient, and finally it has come! It’s the first day of labwork.

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June 19th, 2018: Foundation INSA Toulouse meeting

Looking for funds, we presented our project to the INSA Toulouse foundation.

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June 23rd-24th, 2018: Collaboration with the Montpellier iGEM team

We started a collaboration with the Montpellier team to help them in some areas like the wiki. To do so, we went to Montpellier to talk with them and share our knowledge of the competition.

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June 25th, 2018: TWB meeting

We met Pierre Monsan, one of the main founders of one of our sponsors, Toulouse White Biotechnology (TWB).

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June 26th, 2018: NUS meeting

We organized a Skype meeting with the NUS iGEM team to discuss about a possible collaboration.

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June 27th, 2018: Meeting the STL teachers

As part of our discussions with the french ministry of education, high school teachers visited the lab where we worked during summer.

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June 27th, 2018

For our entrepreneurship approach, we met Guillaume Boissonat who co-funded the startup pili.bio. He shared with us his own experience as a co-funder, and he gave us some advice about the possible business model for a Cerberus startup.

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June 29th, 2018: Primary school intervention

We gave a class in a primary school in order to present Microbiolworld, the research profession, and more generally to introduce the world of microbes to children.

Extra-LabBOOK: JULY

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July 5th, 2018: CALMIP’s meet and visit

We visited CALMIP, a high-throughput calculations center which allowed us to execute complex algorithms for our modelling.

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July 6th, 2018: Official announcement of our project

We officially announced the purpose of our project on social medias!

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July 7th, 2018: 4th Annual parisian meet-up

We took part in the annual parisian meet-up organized by the Pasteur iGEM team.

Meet up Paris
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July 7th-12th, 2018: EuroScience Open Forum (ESOF)

We participated in the EuroScience Open Forum in Toulouse to introduce the iGEM competition and synthetic biology to the public.

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July 19th, 2018: First publications in french newspapers

Newspapers from Toulouse interviewed us about our Cerberus project, we were very happy and grateful for this recognition!

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July 20th-22nd, 2018: European meet-up in Munich

Two members of the team travelled to Munich to represent our team at the European meet-up.

Meet up Munich
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July 30th, 2018: Incubator meeting

We met Yohann Bouvier who is the manager of a startup incubator, named "le starter". He gave us some advice regarding our entrepreneurship approach.

Extra-LabBOOK: AUGUST

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August 1st, 2018: Laval skype

We contacted the Laval iGEM team in order to create an innovative human practices collaboration.

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August 17th, 2018: Photoshoot for the wiki

We dressed up to take the photos for our wiki.

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August 25th-26th, 2018: Collaboration with Bordeaux iGEM team

We went to Bordeaux to bring them our bacterial cellulose, so they could use it as another carbon source for their Far Waste project.

icon

August 29th, 2018: First working group with Le Catalyseur

Le Catalyseur is a business incubator, they helped us to write our business plan. This first session allowed us to structure our company and define our objectives.

icon

August 31st, 2018: Second workshop with Le Catalyseur

During this second meeting, we identified the critical points and risk factors we needed to enhance to conduct Cerberus as a business.

Extra-LabBOOK: SEPTEMBER

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September 4th, 2018

Le Catalyseur helped us to create a business model CANVAS.

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September 5th, 2018:

Le Catalyseur helped us to create a SWOT matrix to define our strengths and weaknesses as a business.

Extra-LabBOOK: OCTOBER

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October 3rd, 2018:

We presented our project in an international high school near Toulouse. This also served as a general repetition for our presentation at the giant Jamboree.

icon

October 5th, 2018:

We discussed industrial property during another meeting with Le Catalyseur.

icon

October 16th, 2018:

We managed to complete all our wiki pages a day earlier, with the precious help of our supervisor Brice. We had to work late into the night, but it was worth it!

wiki pre-freeze picture

No dogs were harmed over the course of this iGEM project.

INSA Toulouse UPS LISBP TWB Defi Adisseo Foundation ESF Catalyses


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