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

From 2014.igem.org

(Difference between revisions)
Line 17: Line 17:
<br>
<br>
<li>Centrifuge 10min at 4000rmp and 4°C
<li>Centrifuge 10min at 4000rmp and 4°C
-
<li>Wash cells 1:1 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O or <u>cold glycerol 10%</u>
+
<li>Wash cells 1:1 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O</u> or <u>cold glycerol 10%</u>
<li>Centrifuge 10min at 4000rmp and 4°C
<li>Centrifuge 10min at 4000rmp and 4°C
-
<li>Wash cells 1:2 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O or <u>cold glycerol 10%</u>
+
<li>Wash cells 1:2 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O</u> or <u>cold glycerol 10%</u>
<li>Centrifuge 10min at 4000rmp and 4°C
<li>Centrifuge 10min at 4000rmp and 4°C
-
<li>Wash cells 1:10 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O or<u> cold glycerol 10%</u>
+
<li>Wash cells 1:10 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O</u> or<u> cold glycerol 10%</u>
<li>Centrifuge 10min at 4000rmp and 4°C
<li>Centrifuge 10min at 4000rmp and 4°C
-
<li>Resuspend cells 1:100 of the initial volume with <u>cold sorbitol</u>, <u>cold MiliQ H2O or <u>cold glycerol 10%</u>
+
<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
<li>Stock cells in aliquots of 50µL at -80°C
</ul>
</ul>

Revision as of 13:08, 4 September 2014

Picture

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

Aug 05