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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;"> | ||
+ | |||
+ | <h2>Nothing done at the bench this week because of material issues</h2> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | |||
+ | <div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week23)"><h3>Week 3 :</h3><img src="aaa" width=600px" height="200px"/></div> | ||
+ | <div id="week3" style="display: none;"> | ||
+ | |||
+ | <u>Tris-EDTA preparation (Sofiane):</u><br> | ||
+ | <ol> | ||
+ | <li>➟10mL 1M Tris pH 7.5 + 2mL 0,5M EDTA pH 8.0</li><br> | ||
+ | <li>➟0,5M EDTA : 18,6g EDTA + 100 mL H2O. Ajuste pH 8.0 with solid NaOH (2,6g)</li> <br> | ||
+ | <li>➟1M Tris : 30,285g + 205 mL H2O. Ajust pH 7.5 with HCl. </li><br> | ||
+ | </ol> | ||
+ | <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> | ||
+ | <br> | ||
+ | <u><b>Competency test (Djamila/Zinebe)</b></u> | ||
+ | <br> | ||
+ | <u>Chloramphenicol (cm 25 mg/mL) preparation: </u><br> | ||
+ | <br> | ||
+ | 0.25 g diluted in 10 mL EtOH 95%; aliquots at -20<br> | ||
+ | <br> | ||
+ | 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> | ||
+ | <br> | ||
+ | <u>Cm (Cf = 5 ug/mL) LB plates:</u><br> | ||
+ | |||
+ | 70 mL melted LB agar + 14 uL cm <br> | ||
+ | <u>Note:</u> agar cooled too much before pouring plate (only got 3 plates) so made more<br> | ||
+ | <br> | ||
+ | 100 mL melted LB agar + 20 uL cm<br> | ||
+ | <br> | ||
+ | <u><b>Competency test (CT) </u></b><br> | ||
+ | <br> | ||
+ | Made SpII-EGTA solution (solutions made yesterday by Sofiane) <br> | ||
+ | <br> | ||
+ | <u>T base 200 ml :</u><br> | ||
+ | <ol> | ||
+ | <li>➟50% (w/v) glucose 2 ml<br> | ||
+ | <li>➟1.2% (w/v) MgSO4·3H2O 14 ml<br> | ||
+ | <li>➟10% (w/v) Bacto yeast extract 2 ml<br> | ||
+ | <li>➟1% (w/v) casamino acids 2 ml<br> | ||
+ | <li>➟4 ml EGTA (0.1 M, pH 8.0) <br> | ||
+ | </ol> | ||
+ | Aliquots put at -20°C<br> | ||
+ | <br> | ||
+ | After thawing competent BSU9716 at 37C in incubator (water bath had issues), added 500 uL of SpII-EGTA and mixed up and down. <br> | ||
+ | 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> | ||
+ | Bacteria+DNA mixture incubated at 37°C with shaking (150 rpm) for an hour <br> | ||
+ | Spreaded the bacteria on Cm LB agar plate then put at 37°C overnight<br> | ||
+ | At this point from the rest of competent bacteria, 100 uL spread on Kan plates <br> | ||
+ | <br> | ||
+ | <u>Note: </u><br> | ||
+ | Should have spread the transformed bacteria on Cm and Kan plates<br> | ||
+ | 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> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week4)"><h3>Week 4 :</h3><img src="aaa" width=600px" height="200px"/></div> | ||
+ | <div id="week4" style="display: none;"> | ||
+ | |||
+ | <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> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week5)"><h3>Week 5 :</h3><img src="aaa" width=600px" height="200px"/></div> | ||
+ | <div id="week5" style="display: none;"> | ||
+ | |||
+ | <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> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week6)"><h3>Week 6 :</h3><img src="aaa" width=600px" height="200px"/></div> | ||
+ | <div id="week6" style="display: none;"> | ||
+ | |||
+ | <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> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week7)"><h3>Week 7 :</h3><img src="aaa" width=600px" height="200px"/></div> | ||
+ | <div id="week7" style="display: none;"> | ||
+ | |||
+ | <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> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div align="center" style="margin: 10px 0px 10px;" onClick="ShowHide(week8)"><h3>Week 8 :</h3><img src="aaa" width=600px" height="200px"/></div> | ||
+ | <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