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 M9 = 0.15 <br>
-
> OD(600nm) in MB =0.4 <br>
+
> OD(600nm) in MB = 1.5 <br>
<br>
<br>
<b>WORK IN ICE</b> <br>
<b>WORK IN ICE</b> <br>

Revision as of 14:28, 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.
    > 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