High Efficiency Transformation 

Thaw a tube of competent E. coli cells on ice for 0h 10m 0s. Mix gently and carefully pipette 50µL of cells into a transformation tube on ice.

Add 1µL containing 100ng 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 0h 30m 0s. Do not mix.

Heat shock at exactly 42°C for exactly 0h 0m 30s. Do not mix.

Place on ice for 0h 5m 0s. Do not mix.

Pipette 950µL of room temperature SOC into the mixture.

Place at 37°C for 1h 0m 0s. Shake vigorously (250rpm) or rotate.

Warm selection plates to 37°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 37°C. Alternatively, incubate at 30°C for 24h 0m 0s or 25°C for 48 hours.