Team:Grenoble-EMSE-LSU/Documentation/Notebook/June

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<h1>June</h1>
<h1>June</h1>
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                                          <h2>Week 1 (3-7)</h2>
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                                  <h2>Week 1 (3-7)</h2>
                                           <h3>Monday</h3>
                                           <h3>Monday</h3>
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<div id="box" style="width: 700px; margin-left: 137px; padding: 5px; border: 3px solid #000; background-color: #fe2b33;"> <div id="template" style="text-align: center; font-weight: bold; font-size: large; color: #f6f6f6; padding: 5px;"> READ THESE INSTRUCTIONS. </div> <div id="instructions" style="text-align: center; font-weight: normal; font-size: small; color: #f6f6f6; padding: 5px;"> Sarah is currently working on this page. <strong>DO NOT EDIT THIS PAGE!!!</strong> </br> </div> </div>
 
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                                               <p>
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  <img src="https://static.igem.org/mediawiki/2013/1/1c/Grenoble_sarah_travaille.jpg"width="600px" />
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  <img src="https://static.igem.org/mediawiki/2013/1/1c/Grenoble_sarah_travaille.jpg"width="300px" />
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</p><p id="legend">Sarah working on the association guide.</p>
                                               <h3>Wednesday</h3>
                                               <h3>Wednesday</h3>
                                                   <p>Beginning of ‘intra-cell ROS concentration’ modelisation.</br>
                                                   <p>Beginning of ‘intra-cell ROS concentration’ modelisation.</br>
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Preparation of competent cells via the TSS method:<br>
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Preparation of competent cells via <a href="https://static.igem.org/mediawiki/2013/f/fe/Grenoble_Protocols-TSS_Transformations.pdf">the TSS method </a><br>
This method gives cells that are more competent than with CaCl2, but the reagents are harder to prepare. The cells can be stored at -80°C like CaCl2-competent cells.<br><br>
This method gives cells that are more competent than with CaCl2, but the reagents are harder to prepare. The cells can be stored at -80°C like CaCl2-competent cells.<br><br>
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2X TSS composition is:<br><br>
 
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10% in volume DMSO
 
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20% in mass PEG-6000 or 8000
 
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MgSO4 at 100 mM concentration<br><br>
 
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in Lysogeny Broth (Luria Broth) at final pH=6.5<br><br>
 
Transfection of competent cells with the biobricks :</br>
Transfection of competent cells with the biobricks :</br>
- pLac</br>
- pLac</br>
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                                                 <p><strong>Construction of the pQE30::KillerRed vector</strong></br>
                                                 <p><strong>Construction of the pQE30::KillerRed vector</strong></br>
Mini-prepped pQE30::&alpha;SNAP/pRep4 and got a 98,9 ng/µL DNA sample.<br/> <br/>
Mini-prepped pQE30::&alpha;SNAP/pRep4 and got a 98,9 ng/µL DNA sample.<br/> <br/>
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Digested pQE30::&alpha;SNAP with BamHI and KpnI restriction enzymes. Separation of the gene of non interest (&alpha;SNAP) from the pQE30 vector backbone by gel electrophoresis (1.2 % agarose, 30 min, 135V).<br/> <br/>
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<a href="https://static.igem.org/mediawiki/2013/e/e3/Grenoble_Digestion_ofpQE30alphaSNAP.pdf">Digested pQE30::&alpha;SNAP with BamHI and KpnI restriction enzymes.</a> Separation of the gene of non interest (&alpha;SNAP) from the pQE30 vector backbone by gel electrophoresis (1.2 % agarose, 30 min, 135V).<br/> <br/>
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Latest revision as of 01:06, 5 October 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM