Team:Evry/Notebook/Protocols/Transformation

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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

    • Retrieved from "http://2014.igem.org/Team:Evry/Notebook/Protocols/Transformation"