IGEM:British Columbia/Protocols/Transformation Competent Cells

Materials

• Competent cells
• Plasmid to be transformed
• Ice
• LB medium
• LB agar plates

Equipment

• Water bath/heat block at 42°C

Method

• Remove 100μL aliquots of competent cells from -80°C freezer and thaw on ice
• Add 1μL of plasmid to be transformed to the thawed cells and incubate for 30 minutes on ice

NOTE: If working from a miniprep plasmid stock, the concentration will likely be too high and will have to be diluted.

• Heat shock in water bath/heat block for 60 seconds
• Put cells back on ice for 2 minutes
• Add 400μL LB medium and incubate at 37°C for 2 hours
• Plate entire 500μL onto a LB agar plate with the correct antibiotic and spread evenly with a sterile spreader
• Incubate overnight (<18hrs) at 37°C