Bacterial Transformation

Protocol

• Thaw 100 μL of competent cells (per transformation) on ice just before they are needed;
• Add DNA (max 20μl) thawed cells and mix by flicking the side of the tube. Leave on ice for 30 minutes;
• Heat shock 5 minutes at 37 degrees Celsius;
• Place on ice for 5 minutes;
• Add 250ul SOC medium to each tube;
• Incubate for 30 to 60 minutes with shaking at 37 degrees Celsius (note that for Kanamycin containing plasmids always use one hour);
• Spin down to remove all supernatant except approximately 100 μL;
• Plate approximately 50 μL on antibiotic plates;
• Grow overnight at 37 degrees Celsius.