Protocols

Transformation of Rubidium Chloride Chemically Competent Cells

1. Thaw cells on ice.
2. Aliquot 100 ml cells per reaction into chilled eppendorf tubes.
3. Add 10 ml DNA per reaction if ligation. Add 1 ml DNA otherwise.
4. Incubate 20 minutes at 4°C (on ice).
5. Heat shock 42°C for 2 minutes and put on ice for 1 minute.
6. Add 1 ml SOC media.
7. Incubate 37°C for 30 minutes.
8. Plate desired amount on appropriate media (LB+AMP)
9. To plate all:
   1. spin 6000rpm for 1 minute
   2. discard supernatant
   3. resuspend in appropriate media
   4. plate on appropriate media.
10. Allow plates to incubate for about 16 hours at 37°C.