Team:Grenoble/Biology/Protocols/Transformation 2


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iGEM Grenoble 2012


Transformation of E. coli competent cells (heat shock)


Transform E. coli competent cells with foreign DNA.


Following are the instructions from the OpenWetWare website.

  1. Thaw TSS cells on ice.
  2. Add DNA, pipette gently to mix (1 µL of prepped plasmid is more than enough).
  3. Let sit for 30 minutes on ice.
  4. Incubate cells for 30 seconds at 42°C.
  5. Incubate cells on ice for 2 min.
  6. Add 1 mL of LB medium at room temperature.
  7. Incubate for 1 hour at 37°C on shaker.
  8. Spread 100-300 µL onto a plate made with appropriate antibiotic.
  9. Grow overnight at 37°C.