From 2007.igem.org

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Revision as of 11:07, 3 August 2007

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.

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