Team:Paris/July 16

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{{Paris/Calendar_Links|July 15|July 17}}
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== Results of transformations ==
== Results of transformations ==
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* Transformations from biobricks 2008 don't succeed '''again'''.
* Transformations from biobricks 2008 don't succeed '''again'''.
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== Preparation of competent cells ==
== Preparation of competent cells ==

Latest revision as of 18:47, 16 July 2008

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Results of transformations

  • Positive controls from biobricks 2007, highest number of bacterias than the first try. It's probably due to the highest quantity of bacteria introduced (150µL instead 100µL).
  • Transformations from biobricks 2008 don't succeed again.

Preparation of competent cells

  • Dilutions 1/200
  • Culture during 3h --> DO= 0,15 / 4h --> DO= 0,13
  • problem from the spectro, culture have been thrown.


Transformations Biobricks 2008

use of a new protocol --> directly competent cells

  • Defroze competent cells on ice during 5'
  • Mix 100µL of competent bacterias with 5µL of biobricks 08 (or 1µL for the positive control)
  • Incubate 10' on ice
  • Heat-shock the cells during 45-50" at 42°C (don't shake!!!)
  • Put 2' on ice
  • Add 900µL of cold SOC medium (4°C)
  • Incubate 1h at 37°C under shaking (225rpm)
  • Transfer in an eppendorf tube and spin at 5000rpm during 30"
  • Remove 800µL of supernatant
  • Resuspent the pellet in the 150-200µL left
  • Spread on plates
  • Incubate O/N at 37°C
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