IGEM:British Columbia/2009/Protocols/Transformation

Transformation (adapted from OWW)

Supplies Needed:

• Competent cells
• Water bath/heat block at 42°C
• Ice
• SOC/LB medium
• LB agar plates (with appropriate selection)
• Plasmid DNA
• Shaking incubator at 37°C

Steps:

1. Remove competent cells (100uL aliquots) from -80°C and thaw on ice.
2. Add 1uL 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.
4. Add 400uL SOC/LB medium and incubate at 37C for 1-2 hours.
5. Plate one plate each of 100uL and 400uL, and spread evenly with sterile glass beads.
6. Incubate overnight (<18hrs) at 37°C.