Difference between revisions of "Team:Toulouse France/Notebook"

Revision as of 17:05, 23 September 2016

iGEM Toulouse 2016

Notebook

Jun.

15
25
Jul.

5
15
25
Aug.

5
15
25
Sept.

5
15
25
Oct.

19

First of all, we had to digest the backbone pSB1C3, which have to received each of our parts. Thus we obtained high quantity of pSB1C3 plasmids with RFP insert due to cultures, and we purified and digested it with the enzymes EcoRI and PstI. As the gBlocks were received, we did PCR to amplify them and create a stock. Most of the PCR worked, but not all of them. Thus, we start trying to clone the good amplicons in the backbone pSB1C3, but it fail because of a lack of genetic material.
We asked to iGEM Munich 2012 to send us four shuttle backbones, that can be used in E. Coli as well as in B. subtilis. We amplified them and stocked them. In parallel, we have done petri dishes with several antibiotic resistances to have it in advance in stock.
In 2014, the iGEM Toulouse team worked on fungi, and constructed strains able to secrete antifungals. We used these strains in order to: have a positive control when our bacteria will be functional, test the efficiency of the antifungals, and find the best temperature, medium and technique. We also determined what was the more efficient between using the surnageant, the bacteria, or a mix of both. All these tests were performed on a fungi mat.

Dry lab: Now that the subject is defined, and that the bibliography is done, let’s prepare our construction! What we have done:
- selected and adapted genes
- designed primers

Wet lab: Having a good control of the basic experiments, as enzymatic digestion or transformation for example, is important to ensure the lab work during the summer. We have done PCR, digestion, ligation, and transformation for clonings during a week, being supervised by PhD students.
Dry lab: In parallel, we finished ordering gBlocks from IDT, and we refined the gene design.

On est motivées

Wiki Freeze !!!!

Website by Team iGEM Toulouse 2016

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 					</div>
-					<div id="shiji_1p">Debut =D.</div>
+					<br>
-					<div id="shiji_2p">On est motivées</div>
+					<div style="margin:0px 10%;">
+						<div class="hoc container clear">
+							<br><br>
+							<div class="group center">
+									<p align="justify" style="font-size:15px;">
+					First of all, we had to digest the backbone pSB1C3, which have to received each of our parts. Thus we obtained high quantity
+					of pSB1C3 plasmids with RFP insert due to cultures, and we purified and digested it with the enzymes EcoRI and PstI. As the
+					gBlocks were received, we did PCR to amplify them and create a stock. Most of the PCR worked, but not all of them. Thus, we
+					start trying to clone the good amplicons in the backbone pSB1C3, but it fail because of a lack of genetic material.
+					<br>
+					We asked to iGEM Munich 2012 to send us four shuttle backbones, that can be used in E. Coli as well as in B. subtilis.
+					We amplified them and stocked them. In parallel, we have done petri dishes with several antibiotic resistances to have
+					it in advance in stock.
+					<br>
+					In 2014, the iGEM Toulouse team worked on fungi, and constructed strains able to secrete antifungals. We used these strains
+					in order to: have a positive control when our bacteria will be functional, test the efficiency of the antifungals, and
+					find the best temperature, medium and technique. We also determined what was the more efficient between using the
+					surnageant, the bacteria, or a mix of both. All these tests were performed on a fungi mat.
+					<br>
+					</p>
+					</div>
+					<div id="shiji_1p">Dry lab: Now that the subject is defined, and that the bibliography is done, let’s prepare our construction!
+					What we have done:
+					<br>
+					- selected and adapted genes <br>
+					- designed primers <br>			</div>
+					<div id="shiji_2p">Wet lab: Having a good control of the basic experiments, as enzymatic digestion or transformation for example,
+					is important to ensure the lab work during the summer. We have done PCR, digestion, ligation, and transformation for clonings
+					during a week, being supervised by PhD students. <br>
+					Dry lab: In parallel, we finished ordering gBlocks from IDT, and we refined the gene design. </div>
 					<div id="shiji_3p">On est motivées</div>
 					<div id="shiji_4p">On est motivées</div>