Difference between revisions of "Team:Paris Bettencourt/Protocols"
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+ | |||
+ | <div class="textBox" id="ElectrocompetentCells"> | ||
+ | <h3>Electrocompetent Cells</h3> | ||
+ | <ul> | ||
+ | <li>Inoculate two 250mL LB flasks with 100µL of an overnight culture of DH5α | ||
+ | <li>Incubate until the the DO600 reach 0.5 to 0.7 | ||
+ | <li>Place the cultures on ice for 15 minutes | ||
+ | <li>For the culture in cold sterile 50mL falcon tubes | ||
+ | <li>Centrifuge them for 10 minutes at 6000rpm | ||
+ | <li>Throw the supernatant | ||
+ | <li>Resuspend the cells in 50mL cold distilled water | ||
+ | <li>Centrifuge them for 10 minutes at 6000rpm | ||
+ | <li>Throw the supernatant | ||
+ | <li>Resuspend the cells in 25mL cold distilled water | ||
+ | <li>Centrifuge them for 10 minutes at 6000rpm | ||
+ | <li>Throw the supernatant | ||
+ | <li>Resuspend the cells in 12.5mL cold 10% glycerol | ||
+ | <li>Centrifuge them for 10 minutes at 6000rpm | ||
+ | <li>Throw the supernatant | ||
+ | <li>Resuspend the cells in 5mL cold 10% glycerol | ||
+ | <li>Make aliquots of the desire volume in microcentrifuge tubes and freeze them at -80°C | ||
+ | </ul> | ||
+ | </div> | ||
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<div class="innerBox yellow"> | <div class="innerBox yellow"> | ||
− | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols | + | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#ElectrocompetentCells"> |
− | <div class="ptext"><p>Electrocompetent Cells | + | <div class="ptext"><p>Electrocompetent Cells</p></div> |
<img src="https://static.igem.org/mediawiki/2015/d/da/ParisBettencourt_protocolsErlen.JPG"> | <img src="https://static.igem.org/mediawiki/2015/d/da/ParisBettencourt_protocolsErlen.JPG"> | ||
</a> | </a> |
Revision as of 14:39, 10 September 2015