Revision as of 14:23, 21 May 2009 by Amelia Hardjasa (New page: ==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) * Plasm...)
Transformation (adapted from OWW)
- Competent cells
- Water bath/heat block at 42°C
- SOC/LB medium
- LB agar plates (with appropriate selection)
- Plasmid DNA
- Shaking incubator at 37°C
- Remove competent cells (100uL aliquots) from -80°C and thaw on ice.
- Add 1uL ligation mix to thawed cells and incubate for 30 minutes on ice.
- Heat shock in water bath/heat block for 60 seconds and put back on ice for two minutes.
- Add 400uL SOC/LB medium and incubate at 37C for 1-2 hours.
- Plate one plate each of 100uL and 400uL, and spread evenly with sterile glass beads.
- Incubate overnight (<18hrs) at 37°C.