Bologna University/Transformations


1. Thaw the competent cells in ice (do not refreeze).
2. Dispense 100 μl of cells into microfuge tubes on ice.
3. Add 0.1-0.3 μg of plasmid DNA or the respective amount of the ligation reaction.
4. Keep on ice for 30 min.
5. Heat at 42 °C for 60 seconds without agitation.
6. Keep on ice for 2 minutes.
7. Add 0.9 ml of LB medium at room temperature.
8. Incubate at 37 °C for 1 hr with agitation.
9. Pellet the cells and discard supernatant except about 100 μl.
10. Streak on plates containing appropriate antibiotics.
11. Incubate the plates overnight at 37 °C.