Wisconsin/Protocol:Transformation

From 2007.igem.org

< Wisconsin
Revision as of 22:16, 5 July 2007 by Klapoetke (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

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 42oC 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 37oC for 1 hour
  8. Spread ??? uL onto plate... centrifuge first???
  9. Incubate plates for 16 hours at 37oC. Put plates upside down to prevent condensation on surface.
Retrieved from "http://2007.igem.org/wiki/index.php/Wisconsin/Protocol:Transformation"