Team:Grenoble/Biology/Protocols/Transformation 1

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Revision as of 19:08, 27 August 2012

iGEM Grenoble 2012

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Project

Transformation of E. coli competent cells (TSS method)

Goal

Transform E. coli competent cells whith foreign DNA.

Protocol

You need to:
  1. Grow a liquid culture overnight.
  2. Dilute this culture at 1/100, in fresh LB medium.
  3. Let the culture grow to 0.3 OD600.
  4. Put 50 µL of the culture in 50 µL of cold TSS 2X.
  5. Incubate cells for 45 min in glaced water.
  6. Add 3 µL of DNA.
  7. Incubate cells for 10 min in glaced water.
  8. Incubate cells for 1.5 min at 42°C.
  9. Incubate cells for 10 min in glaced water.
  10. Add 900 µL of LB medium.
  11. Incubate for 1 hour at 37°C on shaker.

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