Equipment:

- Heating bath

- Incubator

Chemicals & consumables:

- Ice

- Pipets + steril tips

- LB medium

- LB-Agar-Plates + antibiotics

Procedure:

- Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.

- Add the DNA and incubate the suspension for 15 minutes on ice.

- Heat shock is done by incubating the cells for 45 seconds at 42°C. Put samples back on ice for 2 min.

- Add 1 mL of LB medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.

- It might be useful to spin down cells at 5000 rpm for 5 min. Resuspend pellet in 100 µL LB.

Finally the cells are ready to be spread out.