Team:Evry/Notebook/Protocols/Transformation

From 2014.igem.org

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Revision as of 20:08, 17 October 2014

Transformation of Pseudovibrio
  • Place electroporation tanks in ice for 10min


  • Take samples of plasmids (25ng-50ng/µL) /!\ Keep them in ice /!\


  • Take sample of competent cells /!\ Keep them in ice /!\


  • Place 1µL of plasmid in the sample of competent cells


  • Transfer the full volume obtained in the electroporation tank


  • Place in the electroporator and pulse at 2000V

    • NB: The optimal pulse length is between 5 and 6ms.

  • Add 1mL of MB 1X in the 30 seconds following the transformation


  • Incubate between 2h and 3h at 30°C with shaking


  • Centrifuge to concentrate all cells in the pellet


  • Discard the supernatant


  • Sowed the pellet on selective plates of MB 1X


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