Team:Groningen/Template/MODULE/Notebook/protocols/transformationlactococcus

An overnight culture of L. lactis is made in SMGG. The following day, 10 ml of the overnight culture is pipetted in 100 ml SMGG and grown at 30 °C until an OD₆₀₀ of 0.2 - 0.7 is reached. The cells are then three times washed with 50 ml ice cold wash buffer (0.5 M sucrose and 10% glycerol). The cells are then resuspended in 1 ml wash buffer and can be used directly or stored at -80 °C.

For the electroporation 1 μl of the DNA is added to 40 μl cells in an ice cold cuvette. The cuvette is then electroporated at 2.5 kV, 25 μF and 200 Ω. Then, 1 ml SMG17MC is added and the cells are incubated at 30 °C for two hours. The cells can then be plated on GSM17-agar plates.