Bacterial transfomation of plasmid DNA using competent cells

1. Thaw DNA
2. Once DNA is thawed, get out competent cell aliquots from -80 C freezer, thaw on ice
3. Add 1 uL of DNA to cells, flick tube gently to mix
4. Incubate cells on ice for 10 minutes
5. Heat shock at 42 C for 30 seconds
6. Place cells back on ice for 2 minutes
7. Add 1 mL of LB to tubes, shake cells at 37 C for 45 minutes
8. Spin tubes at 4000 rpm for 2 minutes
9. Remove supernatant until about 300 uL remains, gently resuspend cell pellet
10. Pipette suspension onto agar plate with appropriate antibiotic, incubate at 37 C overnight

Bacterial transformation using super-competent cells (use for TOPO, Quik-Change, or ligation DNA samples only)

1. Thaw DNA
2. Once DNA is thawed, get out competent cell aliquots from -80 C freezer, thaw on ice
3. Add 1 uL of DNA to cells, flick tube gently to mix
4. Incubate cells on ice for 30 minutes
5. Heat shock at 42 C for 30 seconds
6. Place cells back on ice for 2 minutes
7. Add 1 mL of LB to tubes, shake cells at 37 C for 60 minutes
8. Spin tubes at 4000 rpm for 2 minutes
9. Remove supernatant until about 300 uL remains, gently resuspend cell pellet
10. Pipette suspension onto agar plate with appropriate antibiotic, incubate at 37 C overnight