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Team:Evry/Notebook/TransformationProto/08-05-2014
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<li>Incubation at 30°C and waiting for the exponential growth phase.<br> | <li>Incubation at 30°C and waiting for the exponential growth phase.<br> | ||
<br> | <br> | ||
| - | <b>WORK IN ICE</b> | + | <b>WORK IN ICE</b> <br> |
<br> | <br> | ||
<li>Centrifuge 10min at 4000rmp and 4°C | <li>Centrifuge 10min at 4000rmp and 4°C | ||
Revision as of 11:39, 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.
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
