From 2007.igem.org

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. Transfer cells to new culture tubes (more O₂ for growth) and add 900uL of SOC to each culture tube
7. Incubate on shaker at 37^oC for 1 hour
8. Grow 4 plates: 1 normal diluted, 1 normal concentrated, and 2 controls concentrated.
   • Diluted: Spread 200uL onto plate
   • Concentrated: Centrifuge all tubes for 5 minutes at 5000rpm first, then pippet everything except for 200uL onto plates.
9. Incubate plates for 16 hours at 37^oC. Plates should face down to prevent condensation on surface.