Team:USTC CHINA/Notebook/Protocols/The transformation of E.coli

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Latest revision as of 08:59, 27 September 2013

The transformation of E.coli

1. Remove competent cells (200μl/tube) from freezer and allow to thaw on ice for 3 min.
2. Add 3-5 μl of DNA to the cells.
3. Incubate on ice for 30 min.
4. Heat shock the cells at 42°C for 45 seconds.
5. Add 800 μl of LB Broth, and incubate in a shaker at 37°C for 45~60min.
6. Centrifuge at 12,000 for 1 min, discard 800 μl supernatant.
7. Re-suspend the pellet in the 200 μl of supernatant and spread onto one agar plate.
8. Incubate the plates overnight at 37°C.

Notebook

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