Transformation Protocol

From 2006.igem.org

1. Thaw the compentent cells by warming in between hands. Cells should be used immediately after being thawed.
2. Add 1μg of DNA immediately to the tube containing the cells.
3. Gently swirl to mix.
4. Place DNA on ice for 10 minutes.
5. Heat shock cells by placing into a 42°C water bath for two minutes.
6. Add 1mL LB no amp medium to each tube.
7. Incubate for one hour at 37°C.
8. Spin cells for 2 min and resuspend the cells in 100μl of LBamp medium.
9. Place μl of transformation on LB/ampicillin-containing plates.
10. Spread over the surface of the plate until dry.
11. Once the plate is dry incubate for 12-16 hours at 37°C.
12. Any remaining transformation mixture can be stored at 4°C and used for subsequent plating.