From 2007.igem.org

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.

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