DNA transformation protocol
- Thaw all reagents completely on ice.
- Add 1 µL of ligation reaction to thawed competent cells.
- Gently mix by tapping tube of competent cells.
- Incubate reaction on ice for 30 minutes.
- Heat shock the competent cell mixture by incubation for 30 to 60 seconds in a 42°C water bath.
- Incubate tubes on ice for 2 minutes.
- Add 250 to 500 µL of SOC or LB media.
- Incubate at 37°C and shake at 250 rpm.
- Warm selection plates to 37°C.
- Spread 10, 50, and 100 µL of transformed cells on selection plates.
- Incubate plates at 30°C overnight.
to 50 µL
SOC or LB media