DNA Transformation Protocol

PCR

Ligation

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.

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