Team:British Columbia/Notebook/Protocols/transformation




Competent cells in 100μL aliquots

Water bath/heat block at 42°C


LB Broth

LB agar plates (with appropriate selection)


1. Remove competent cells (100uL aliquots) from -80°C and thaw on ice.

  • Note, aliquots can be thawed GENTLY by hand if time is an issue*

2. Add 1µL ligation mix to thawed cells and incubate for 30 minutes on ice.

3. Heat shock in water bath/heat block for 60 seconds and put back on ice for two minutes.

  • Remember to turn off the water bath*
  • The following steps must be done near a flame*
  • Don’t forget a growth control, especially if you aren’t screening using antibiotics*

4. Add 400uL LB Broth and incubate at 37C for 2 hours.

  • Note, 1 hour is also sufficient*

5. Spread plate entire 500µL.

  • Turn off the flame*

6. Incubate overnight (<18hrs) at 37°C.