From 2010.igem.org

Transformation

Materials:

- competent cells

- LB 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 37 °C

- shaking incubator at 37 °C.

Protocol:

1. Add 50-100 ng DNA into a 30 μL competent E.coli, and mix gently. Do not mix by pipetting up and down!

2. Incubate tube vial on ice for 15 minutes.

3. Heat-shocks the cells for exactly 5 minutes at 37 °C without shaking.

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

5. Add 800 μL of room temperature LB medium.

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

7. Spin tube (2,000 rpm, 4 minutes), discard supernatant to leave no more than 100 μL, mix and plate on an agar plate.

8. Incubate plates overnight at 37 °C.