Difference between revisions of "Smolke:Protocols/Heat Shock Transformation"

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**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)
 
**1-10 uL ligation product
 
**1-10 uL ligation product
**1 mL SOC (optional)
+
**250 uL SOC
 
**1 LB plate with appropriate antibiotics
 
**1 LB plate with appropriate antibiotics
 
*Procedure
 
*Procedure
 
**Add DNA to cells
 
**Add DNA to cells
 
**Incubate on ice for 0-20 minutes
 
**Incubate on ice for 0-20 minutes
**Heat shock at 42C for 30 seconds
+
**Heat shock at 42 °C for 30 seconds
**Add SOC (optional)
+
**Incubate on ice for 2 minutes
**Incubate on rotator in 37C incubator for 0-60 minutes
+
**Add SOC
**Add to plate
+
**Incubate on rotator in 37 °C incubator for 0-60 minutes
 +
**Plate 50-250 uL

Revision as of 17:29, 15 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
    • 250 uL SOC
    • 1 LB plate with appropriate antibiotics
  • Procedure
    • Add DNA to cells
    • Incubate on ice for 0-20 minutes
    • Heat shock at 42 °C for 30 seconds
    • Incubate on ice for 2 minutes
    • Add SOC
    • Incubate on rotator in 37 °C incubator for 0-60 minutes
    • Plate 50-250 uL