From 2007.igem.org

Transforming Chemically Competent Cells

  1. Thaw competent cells on ice
  2. Chill three 1.5mL tubes in ice bucket. One for transformation and the other two for control.
    • Normal: pippet 100uL of cells and 9uL of ligation reaction into tube
    • Positive Control: pippet 100uL of cells and 2uL of pUC18 or pUC19 into tube
    • Negative Control: pippet 100uL of cells
  3. Sit on ice for 30 minutes
  4. Heat shock in 42oC water bath for 90 seconds
  5. Incubate on ice for 2 minutes
  6. Add 200uL of SOC to each culture tube
  7. Incubate on shaker (nutator) at 37oC for 1 hour
  8. Grow 4 plates: 1 normal diluted, 1 normal concentrated, and 2 controls concentrated.
    • Diluted: Spread 50-100uL onto plate
    • Concentrated: Centrifuge tube for 2-4 minutes at 5000rpm first, then spread everything except for 50-100uL onto plates.
  9. Incubate plates for 16 hours at 37oC. Plates should face down to prevent condensation on surface.