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Team:Evry/Notebook/TransformationProto/08-05-2014
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(Difference between revisions)
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<li>Re-launch of 50mL of Pseudovribrio in MB and M9 from pre-culture of 4th August. <br> | <li>Re-launch of 50mL of Pseudovribrio in MB and M9 from pre-culture of 4th August. <br> | ||
<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> | ||
| + | -> OD(600nm) in M9 =0.1 <br> | ||
| + | -> OD(600nm) in MB =0.4 <br> | ||
<br> | <br> | ||
<b>WORK IN ICE</b> <br> | <b>WORK IN ICE</b> <br> | ||
Revision as of 13:33, 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 =0.1
-> 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
Aug 05
