Koeris/Notebook/2007-1-17: Difference between revisions
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#Add 1ml SOC to 100ul competent cells | #Add 1ml SOC to 100ul competent cells | ||
#Incubate at 37 deg C for 1h at 300rpm | #Incubate at 37 deg C for 1h at 300rpm | ||
#Spread plate 550ul on LB agar with the appropriate resistance markers | |||
Revision as of 11:46, 17 January 2007
Transformations
Standard heat shock protocol usable for any kind of origin. Also good for multi-plasmid transformations
Reagents necessary
- Chemically competent cells (see here)
- Plasmid prep >5ng/ul concentration as the lower limit is acceptable
- SOC
Protocol
- Thaw competent cells on ice
- Add plasmid DNA to cell, stir with pipette tip, do NOT pipette up and down
- Incubate for 30min on ice
- Heat shock at 42 deg C for 30s
- Cool on ice for 5min
- Add 1ml SOC to 100ul competent cells
- Incubate at 37 deg C for 1h at 300rpm
- Spread plate 550ul on LB agar with the appropriate resistance markers
