Team:Evry/Notebook/Protocols/Transformation

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  <li>Place electroporation tanks in ice for 10min</li><br>

  <li>Place electroporation tanks in ice for 10min</li><br>

  <li>Take samples of plasmids (25ng-50ng/µL)    /!\ Keep them in ice /!\ </li><br>

  <li>Take samples of plasmids (25ng-50ng/µL)    /!\ Keep them in ice /!\ </li><br>

  <li>Take sample of competent cells  /!\ Keep them in ice /!\ </li><br>

  <li>Take sample of competent cells  /!\ Keep them in ice /!\ </li><br>

  <li>Place 1µL of plasmid in the sample of competent cells</li> <br>

  <li>Place 1µL of plasmid in the sample of competent cells</li> <br>

  <li>Transfer the full volume obtained in the electroporation tank</li> <br>

  <li>Transfer the full volume obtained in the electroporation tank</li> <br>

  <li>Place in the electroporator and pulse at 2000V </li><br>

  <li>Place in the electroporator and pulse at 2000V </li><br>

  <b>NB:</b>  The optimal pulse length is between 5 and 6ms.</li> <br>

  <b>NB:</b>  The optimal pulse length is between 5 and 6ms.</li> <br>

  <li>Add 1mL of MB 1X in the 30 seconds following the transformation</li> <br>

  <li>Add 1mL of MB 1X in the 30 seconds following the transformation</li> <br>

  <li>Incubate between 2h and 3h at 30°C with shaking</li> <br>

  <li>Incubate between 2h and 3h at 30°C with shaking</li> <br>

  <li>Centrifuge to concentrate all cells in the pellet</li> <br>

  <li>Centrifuge to concentrate all cells in the pellet</li> <br>

  <li>Discard the supernatant</li><br>

  <li>Discard the supernatant</li><br>

  <li>Sowed the pellet on selective plates of MB 1X </li><br>

  <li>Sowed the pellet on selective plates of MB 1X </li><br>

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