Chemical Transformation

See previous protocol: Chemical Competent E. coli Cell Protocol for details on making chemically competent E. coli cells.

Chemical Transformation protocol.

1. Thaw out the 50 µL cell suspension aliquot 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 incubate on ice for 30 minutes.
4. Induce heat shock by incubating cells at 42^oC for 30 seconds.
5. Incubate on ice for 5 minutes.
6. Add 450 µL of LB and incubate at 37^oC for 1 hour.
7. Plate out 50 µL of straight transformant cells & also carry out a 10-fold dilution (plate out 50µL). Note that the plate should contain the appropriate antibiotic at the MBC in order to select for transformants which contain the desired plasmid.
8. Incubate overnnight at 37^oC in a static incubator.