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Team:Evry/Notebook/TransformationProto/08-05-2014
From 2014.igem.org
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Revision as of 14:33, 4 September 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 = 0.15
> OD(600nm) in MB = 1.5
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
