Team:Aix-Marseille/Experiments/Protocols

Preparation of competent bacteria cells

1. Grow a culture of bacteria in LB medium until it reaches OD600 = 0.5.

2. Centrifuge cells for 10 minutes at 3500g at 4°C

3. Resuspend the pellet slowly in 80mL of Tfb1 buffer.

4. Centrifuge 5 min at 3500g at 4°C.

5. Resuspend the pellet in 8 mL of Tbf2 buffer.

6. Incubate 15 min in ice.

7. Aliquot 200µL of cell suspension in sterile eppendorf tubes and conserve it at -80°C.