Wisconsin/Protocol:Transformation
From 2007.igem.org
Transforming Chemically Competent Cells
- Thaw cells on ice
- 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
- Sit on ice for 30 minutes
- Heat shock in 42oC water bath for 30 seconds
- Incubate on ice for 2 minutes
- Transfer cells to new culture tubes (more O2 for growth) and add 900uL of SOC to each culture tube
- Incubate on shaker at 37oC for 1 hour
- Spread ??? uL onto plate... centrifuge first???
- Incubate plates for 16 hours at 37oC. Put plates upside down to prevent condensation on surface.