Team:LMU-Munich/Notebook/Protocols/19 Transformation SSS


Transformation of Chemically Competent Cells

(Source: Sina Science Sevices)

1. Thaw competent cells on ice.

2. To 50µl-220µl of cells, add 5-50ng of plasmid DNA or up to 10µl of ligation reaction. Mix by tapping gently.

3. Incubate the vial(s) on ice for 30 minutes.

4. Heat shock for 45 seconds at 42°C. Do not mix or shake.

5. Chill tubes on ice for about 2 minutes.

6. IMPORTANT: Spin down cells (e.g. for 14-30 seconds at 6000g) and remove supernatant.

7. Resuspend cells in 200µl LB or SOC medium.

8. Shake vial(s) at 37°C for 1 hour.

9. Plate up to 200µl on selective LB agar plates.