2. Cell transformation

From 2008.igem.org

1. Remove vial of pre-prepared TOP10 chemically competent cells from -80˚C freezer on 5th floor then place in ice bucket to allow melting (will melt on ice).
2. Add between 10-100ng of DNA solution (if comes from DNA extraction, estimation may be necessary).
3. Mix the solution gently by tapping – anything more vigorous will result in potential damage to the cells.
4. Incubate on ice for 30 minutes, meanwhile ensure that a water bath is set to 42˚C and pre-heat a vial of SOC medium.
5. Once incubation on ice is completed, transfer the transformation tube to the 42˚C water bath for exactly 30 seconds.
6. Remove the transformation tube from the water bath and add 250µL of SOC medium, then rapidly place on ice for 1 min while being transported to a 37˚C incubator room with a shaker.
7. Shake at 37˚C for 1 hour.
8. Plate on appropriate media and antibiotics overnight, then check for colonies.