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

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.
  > OD(600nm) in M9 =0.1
  > OD(600nm) in MB =0.4
  
  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