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From 2013.igem.org

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*

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