Team:Birkbeck/Chemical Transformation

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Chemical Transformation

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

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.

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