Bologna University/Transformation


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