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

(Difference between revisions)

Revision as of 15:23, 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

Aug 05

@@ Line 10: / Line 10: @@
 <u> <b> Tests for development of the electroporation's protocol </b> </u> <br>
-<ul>
 <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>
@@ Line 27: / Line 27: @@
 <li>Resuspend cells 1:100 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O</u> or <u>cold glycerol 10%</u>
 <li>Stock cells in aliquots of 50µL at -80°C
-</ul>