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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                                               <p>
                                               <p>
                                               We observed that absorbance measurements differed according to the device used to make them. For example, the Tristar microplate reader and the Thermo Electron Corporation Genesys 6 spectrophotometer we were using  gave different OD600 readings for the same samples. This can be explained by the fact that OD600 measurement values come from diffusion rather than actual absorbance, and thus the distance from the sample to the detector is a factor in the reading. Since it is likely these distances are different from one machine to the other, it would explain the differences in measurement.<br><br>
                                               We observed that absorbance measurements differed according to the device used to make them. For example, the Tristar microplate reader and the Thermo Electron Corporation Genesys 6 spectrophotometer we were using  gave different OD600 readings for the same samples. This can be explained by the fact that OD600 measurement values come from diffusion rather than actual absorbance, and thus the distance from the sample to the detector is a factor in the reading. Since it is likely these distances are different from one machine to the other, it would explain the differences in measurement.<br><br>
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                                                   <p>Preparation of freezable competent cells (M15): 40 alicos of 100µL</br>
                                                   <p>Preparation of freezable competent cells (M15): 40 alicos of 100µL</br>
Test of the competent cells.</p>
Test of the competent cells.</p>
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<p>Redaction of a guide for the future grenoble iGEMers, to explain how to manage the association
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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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</p>

Latest revision as of 01:06, 5 October 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM