Bologna University/Transformations
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< Bologna University
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- 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.
