Transformation of plasmids into electrocompetent cells

For more details about making electrocompetent cells, please see our Making electrocompetent E.coli cells protocol.

Electroporation protocol.

  • 1. Thaw out the 50 µL cell suspension aliquot on ice for ca. 30 minutes. Concurrently, cool electroporation tubes on ice.
  • 2. Add 100pg-1µg of plasmid DNA to the cell suspension (approximately 1-5 µL of plasmid solution).
  • 3. Mix plasmid and cells thoroughly and continue to incubate on ice for 5-10 minutes.
  • 4. Transfer the mixture of cells and plasmids to an electroporation tube.
  • 5. Electroporate cells; for E.coli, this is done at 2.5 kV, 200 Ohms, 25 uF.
  • 6. Immediately add 450-950 uL of LB broth.
  • 7. Incubate tubes at 37oC for 60-90 minutes in a statis incubator.
  • 8. Plate 100 uL of cells onto agar plates with appropriate antibiotic resistance.
  • 9. Incubate overnnight at 37oC in a static incubator.

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