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
Aug 05