From 2010.igem.org

Transformation

Materials:

- competent cells

- SOC medium (warmed to room temperature)

- Plasmid DNA or DNA ligation mix

- LB agar plates containing 15-100 μg/mL antibiotic of choice, pre-warmed to 37 °C

- water bath at 42 °C

- shaking incubator at 37 °C.

Protocol:

1. Add 50-100 ng DNA into a 15 μL competent E.coli, and mix gently. Do not mix by pipetting up and down!

2. Incubate tube vial on ice for 30 minutes.

3. Heat-shocks the cells for exactly 30 seconds at 42 °C without shaking.

4. Immediately transfer the tubes back to ice for 2 minutes.

5. Add 250 μL of room temperature SOC medium.

6. Cap tube tightly and shake tube horizontally (225 rpm) at 37 °C for 1 hour.

7. Plate from each tube 100 μL on an agar plate containing antibiotic.

8. Incubate plates overnight at 37 °C.