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

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<u> <b> Tests for development of the electroporation's protocol </b> </u> <br>
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<u> <b><big><FONT COLOR=#003333> Tests for development of the electroporation's protocol</font> </big></b> </u> <br>
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<ul>
 
<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 = 0.15 <br>
 +
> OD(600nm) in MB = 1.5 <br>
<br>
<br>
<b>WORK IN ICE</b> <br>
<b>WORK IN ICE</b> <br>
<br>
<br>
<li>Centrifuge 10min at 4000rmp and 4°C
<li>Centrifuge 10min at 4000rmp and 4°C
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<li>Wash cells 1:1 of the initial volume with cold sorbitol, cold MiliQ H2O or cold glycerol 10%
+
<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 cold sorbitol, cold MiliQ H2O or cold glycerol 10%
+
<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 cold sorbitol, cold MiliQ H2O or cold glycerol 10%
+
<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 cold sorbitol, cold MiliQ H2O or cold glycerol 10%
+
<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>
+
 

Latest revision as of 15:08, 12 October 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

    • Retrieved from "http://2014.igem.org/Team:Evry/Notebook/TransformationProto/08-05-2014"