IGEM:British Columbia/Protocols/Transformation Competent Cells
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Materials
- Competent cells
- Plasmid to be transformed
- Ice
- LB medium
- LB agar plates
Equipment
- Water bath/heat block at 42°C
Method
- Remove 100μL aliquots of competent cells from -80°C freezer and thaw on ice
- Add 1μL of plasmid to be transformed to the thawed cells and incubate for 30 minutes on ice
NOTE: If working from a miniprep plasmid stock, the concentration will likely be too high and will have to be diluted.
- Heat shock in water bath/heat block for 60 seconds
- Put cells back on ice for 2 minutes
- Add 400μL LB medium and incubate at 37°C for 2 hours
- Plate entire 500μL onto a LB agar plate with the correct antibiotic and spread evenly with a sterile spreader
- Incubate overnight (<18hrs) at 37°C