Team:Evry/Notebook/TransformationProto/08-05-2014

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Revision as of 11:38, 4 September 2014

Picture

Tests for development of the electroporation's protocol

  • Re-launch of 50mL of Pseudovribrio in MB and M9 from pre-culture of 4th August.
  • Incubation at 30°C and waiting for the exponential growth phase.

    WORK IN ICE
  • Centrifuge 10min at 4000rmp and 4°C
  • Wash cells 1:1 of the initial volume with cold sorbitol, cold MiliQ H2O or cold glycerol 10%
  • Centrifuge 10min at 4000rmp and 4°C
  • Wash cells 1:2 of the initial volume with cold sorbitol, cold MiliQ H2O or cold glycerol 10%
  • Centrifuge 10min at 4000rmp and 4°C
  • Wash cells 1:10 of the initial volume with cold sorbitol, cold MiliQ H2O or cold glycerol 10%
  • Centrifuge 10min at 4000rmp and 4°C
  • Resuspend cells 1:100 of the initial volume with cold sorbitol, cold MiliQ H2O or cold glycerol 10%
  • Stock cells in aliquots of 50µL at -80°C

Aug 05