2. Cell transformation

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.