Transformation Protocol (C3032)
- Thaw a tube of SHuffle Competent E. coli cells on ice until the last crystals disappear. Mix gently and carefully pipette 50 µl of cells into a transformation tube on ice.
- Add 1–5 µl containing 1 pg–100 ng of plasmid DNA to the cell mixture. Carefully flick the tube 4–5 times to mix cells and DNA. Do not vortex.
- Place the mixture on ice for 30 minutes. Do not mix.
- Heat shock at exactly 42°C for exactly 30 seconds. Do not mix.
- Place on ice for 5 minutes. Do not mix.
- Pipette 950 µl of room temperature SOC into the mixture.
- Place at 30°C for 60 minutes. Shake vigorously (250 rpm) or rotate.
- Warm selection plates to 30°C.
- Mix the cells thoroughly by flicking the tube and inverting, then perform several 10-fold serial dilutions in SOC.
- Spread 50–100 µl of each dilution onto a selection plate and incubate overnight at 30°C. Alternatively, incubate at 25°C for 48 hours.