Bacterial transformation with plasmid
Material
- E. coli competent cell (-80C)
See Preparation of E. coli competent cell. - Sol III in DNA Ligation Kit (TaKaRa 6022, -20C)
- LB plate with antibiotic (4C)
See Plasmid.
Protocol
Vol of competent cell should be more than 10 times of plasmid sol.
- Dissolve competent cell on ice.
- Add 1 ul sol III WITHOUT PIPETTING.
- Add plasmid sol WITHOUT PIPETTING.
- Incubate on ice for 20 min.
- Heat shock at 42C for 1 min.
- Icubate on ice for 5 min.
- Seed on LB plate.
- Incubate at 37C for o/n.