Smolke:Protocols/Heat Shock Transformation: Difference between revisions

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*For each sample
*For each sample
**1 tube of thawed chemically competent cells (keep covered in ice at all times)
**1 tube of thawed chemically competent cells (keep covered in ice at all times)
**1-10uL ligation product
**1-10 uL ligation product
**1 mL SOC (optional)
**1 mL SOC (optional)
**1 LB plate with appropriate antibiotics
**1 LB plate with appropriate antibiotics

Revision as of 17:21, 11 July 2012

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  • For each sample
    • 1 tube of thawed chemically competent cells (keep covered in ice at all times)
    • 1-10 uL ligation product
    • 1 mL SOC (optional)
    • 1 LB plate with appropriate antibiotics
  • Procedure
    • Add DNA to cells
    • Incubate on ice for 0-20 minutes
    • Heat shock at 42C for 30 seconds
    • Add SOC (optional)
    • Incubate on rotator in 37C incubator for 0-60 minutes
    • Add to plate
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