Transformation

Supplies:

• Competent cells in 100μL aliquots.
• Water bath/heat block at 42°C.
• Ice
• LB Broth
• LB agar plates (with appropriate selection)

Steps:

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