Team:Heidelberg/Sandbox20

Preparation:

1. Pipet 1 µl of the plasmid solution into a pre-cooled 1.5 ml reaction tube. Ligation products have to be diluted in H2O (1:4 usually works) or purified in advance.
2. Heat 1 ml of SOC to 37 °C.
3. Pre-cool an 1 mm electroporation cuvette on ice.
4. Thaw an aliquot of electrocompetent cells on ice.

Electroporation Procedure:

1. Pipet 25 µl of the competent cells into the tube with the plasmid solution.
2. Transfer the mixture carefully into the cuvette (avoid air bubbles)

</li>Electroporate at 1800 kV for 5 ms.

1. Immediately add 1 ml SOC, mix by pipetting up and down, and transfer the solution into a 1.5 ml reaction tube
2. Incubate for 1 h at 37 °C at 350 rpm

</li>Plate 100 µl on agar with the respective antibiotics. Note: If more than one plasmid has to be transformed, the procedure stays the same. Both plasmids have to be mixed in equal amount prior to the addition to the cells. Again, only 1 µl should be used. For the last step, agar plates with 2 antibiotics have to be used. }}|