Difference between revisions of "Team:UPMC-Paris/Notebook"

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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week2)"><h3>Week 2 :</h3><img src="aaa" width=600px" height="200px"/></div>
 
<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week2)"><h3>Week 2 :</h3><img src="aaa" width=600px" height="200px"/></div>
 
<div id="week2" style="display: none;">
 
<div id="week2" style="display: none;">
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<h2>Nothing done at the bench this week because of material issues</h2>
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</div>
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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week23)"><h3>Week 3 :</h3><img src="aaa" width=600px" height="200px"/></div>
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<div id="week3" style="display: none;">
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<u>Tris-EDTA preparation (Sofiane):</u><br>
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<ol>
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<li>➟10mL 1M Tris pH 7.5 + 2mL 0,5M EDTA pH 8.0</li><br>
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<li>➟0,5M EDTA : 18,6g EDTA + 100 mL H2O. Ajuste pH 8.0 with solid NaOH (2,6g)</li> <br>
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<li>➟1M Tris : 30,285g + 205 mL H2O. Ajust pH 7.5 with HCl. </li><br>
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</ol>
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<i>Note :</i> pH ajustment failed, I add 25mL of 12% HCl and 12 mL of 20% HCl to have a pH 7,6. <br> Concentration is not good anymore. We have to make this solution again. Maybe buy TE Buffer? Verification of TE we have is needed! <br>
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<br>
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<u><b>Competency test (Djamila/Zinebe)</b></u>
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<br>
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<u>Chloramphenicol (cm 25 mg/mL) preparation: </u><br>
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<br>
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0.25 g diluted in 10 mL EtOH 95%; aliquots at -20<br>
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<br>
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Note: first dilution in milliQ water by mistake; got rid of water and tried to let it dry but without success so will probably just add directly EtOH in to make more Cm <br>
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<br>
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<u>Cm (Cf = 5 ug/mL) LB plates:</u><br>
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70 mL melted LB agar + 14 uL cm <br>
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<u>Note:</u> agar cooled too much  before pouring plate (only got 3 plates) so made more<br>
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<br>
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100 mL melted LB agar + 20 uL cm<br>
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<br>
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<u><b>Competency test (CT) </u></b><br>
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<br>
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Made SpII-EGTA solution (solutions made yesterday by Sofiane) <br>
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<br>
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<u>T base 200 ml :</u><br>
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<ol>
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<li>➟50% (w/v) glucose 2 ml<br>
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<li>➟1.2% (w/v) MgSO4·3H2O 14 ml<br>
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<li>➟10% (w/v) Bacto yeast extract 2 ml<br>
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<li>➟1% (w/v) casamino acids 2 ml<br>
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<li>➟4 ml EGTA (0.1 M, pH 8.0) <br>
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</ol>
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Aliquots put at -20°C<br>
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<br>
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After thawing competent BSU9716 at 37C in incubator (water bath had issues), added 500 uL of SpII-EGTA and mixed up and down. <br>
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Added 50 uL bacteria in 5 eppendorf tubes after spinning CT plasmids, added 1 uL of DNA in the appropriate tube (50pg/ul, 20pg/ul, 10pg/ul, 5pg/ul, 0.5pg/ul). <br>
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Bacteria+DNA mixture incubated at 37°C with shaking (150 rpm) for an hour <br>
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Spreaded the bacteria on Cm LB agar plate then put at 37°C overnight<br>
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At this point from the rest of competent bacteria, 100 uL spread on Kan plates <br>
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<br>
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<u>Note: </u><br>
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Should have spread the transformed bacteria on Cm and Kan plates<br>
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Competency test kit contains five vials of purified DNA from BBa_J04450 (RFP construct) in plasmid backbone pSB1C3 (cm resistant). Each vial contains DNA at a different concentration: 50pg/ul, 20pg/ul, 10pg/ul, 5pg/ul, 0.5pg/ul. <br>
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</div>
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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week4)"><h3>Week 4 :</h3><img src="aaa" width=600px" height="200px"/></div>
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<div id="week4" style="display: none;">
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<h2>Ca me soule toujours autant</h2>
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<ol><li>Benjamin : Il s'est fait chier</li>
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<li>Nassim : Il a cassé les couilles</li>
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<li>Sofiane : Il a tout fait cramer</li>
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<li>Wilhelm : Il est reparti</li>
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<li>Kévin : Il est pas venu</li>
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<li>Perwin : Continuation de la quête chez les RG</li>
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<li>Zinebe : Elle a volé le PLA</li>
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<li>Djamila : Comme d'hab, le quota</li>
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<li>Les autres : Vacances</li></ol>
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</div>
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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week5)"><h3>Week 5 :</h3><img src="aaa" width=600px" height="200px"/></div>
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<div id="week5" style="display: none;">
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<h2>Ca me soule toujours autant</h2>
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<ol><li>Benjamin : Il s'est fait chier</li>
 +
 +
<li>Nassim : Il a cassé les couilles</li>
 +
 +
<li>Sofiane : Il a tout fait cramer</li>
 +
 +
<li>Wilhelm : Il est reparti</li>
 +
 +
<li>Kévin : Il est pas venu</li>
 +
 +
<li>Perwin : Continuation de la quête chez les RG</li>
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 +
<li>Zinebe : Elle a volé le PLA</li>
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<li>Djamila : Comme d'hab, le quota</li>
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<li>Les autres : Vacances</li></ol>
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</div>
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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week6)"><h3>Week 6 :</h3><img src="aaa" width=600px" height="200px"/></div>
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<div id="week6" style="display: none;">
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<h2>Ca me soule toujours autant</h2>
 +
 +
<ol><li>Benjamin : Il s'est fait chier</li>
 +
 +
<li>Nassim : Il a cassé les couilles</li>
 +
 +
<li>Sofiane : Il a tout fait cramer</li>
 +
 +
<li>Wilhelm : Il est reparti</li>
 +
 +
<li>Kévin : Il est pas venu</li>
 +
 +
<li>Perwin : Continuation de la quête chez les RG</li>
 +
 +
<li>Zinebe : Elle a volé le PLA</li>
 +
 +
<li>Djamila : Comme d'hab, le quota</li>
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<li>Les autres : Vacances</li></ol>
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</div>
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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week7)"><h3>Week 7 :</h3><img src="aaa" width=600px" height="200px"/></div>
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<div id="week7" style="display: none;">
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<h2>Ca me soule toujours autant</h2>
 +
 +
<ol><li>Benjamin : Il s'est fait chier</li>
 +
 +
<li>Nassim : Il a cassé les couilles</li>
 +
 +
<li>Sofiane : Il a tout fait cramer</li>
 +
 +
<li>Wilhelm : Il est reparti</li>
 +
 +
<li>Kévin : Il est pas venu</li>
 +
 +
<li>Perwin : Continuation de la quête chez les RG</li>
 +
 +
<li>Zinebe : Elle a volé le PLA</li>
 +
 +
<li>Djamila : Comme d'hab, le quota</li>
 +
 +
<li>Les autres : Vacances</li></ol>
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</div>
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<div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week8)"><h3>Week 8 :</h3><img src="aaa" width=600px" height="200px"/></div>
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<div id="week8" style="display: none;">
  
 
<h2>Ca me soule toujours autant</h2>
 
<h2>Ca me soule toujours autant</h2>

Revision as of 12:13, 18 October 2016