Transformation Protocol.  
Thaw competent cells on ice Chill approximately 5 ng (2 μl) of the ligation mixture in a 1.5 ml microcentrifuge tube.
Add 50 µl of competent cells to the DNA.
Mix gently by pipetting up and down or flicking the tube 4–5 times to mix the cells and DNA.
Do not vortex.
Place the mixture on ice for 30 minutes.
Do not mix.
Heat shock at 42°C for 30 seconds.
Do not mix.
Add 950 µl of room temperature media to the tube.
Place tube at 37°C for 60 minutes.
Shake vigorously (250 rpm) or rotate.
Warm selection plates to 37°C.
Spread 50–100 µl of the cells and ligation mixture onto the plates.
Incubate overnight at 37°C
