Transformation of E. coli

Transformation of E. coli

  • Thaw competent cells carefully on ice
  • Incubate the DNA with the competent cells for 1-10 min on ice
  • Incubate for 30-90 sec. at 42°C - heat shock -
  • Spread the cells on the plates with the appropriate selection marker
  • Incubate plates over night at 37°C

This procedure works excellent with ampicillin as a selection marker.

With kanamycin or streptomycin transformation rates are ten times lower.

With chloramphenicol as a selection marker this method failed in our hands

For more information see:

Brian Pope and Helen M. Kent
High efficiency 5 min transformation of Escherichia coli
Nucleic Acids Research 1996, Vol. 24, No. 3, 536-537