From 2007.igem.org

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Revision as of 22:16, 5 July 2007

Transforming Chemically Competent Cells

1. Thaw cells on ice
2. Pippet 100uL of cells into three 1.5mL tube. One for transformation and the other two for control.
   • Normal: pippet 9uL of DNA into tube
   • Positive Control: pippet 2uL of pUC18 or pUC19 into tube
   • Negative Control: nothing
3. Sit on ice for 30 minutes
4. Heat shock in 42^oC water bath for 30 seconds
5. Incubate on ice for 2 minutes
6. Add 900uL of SOC to each tube
7. Incubate on shaker at 37^oC for 1 hour
8. Spread ??? uL onto plate... centrifuge first???
9. Incubate plates for 16 hours at 37^oC. Put plates upside down to prevent condensation on surface.