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Team:Montreal/Cell transformation
From 2008.igem.org
(New page: 1. Remove vial of pre-prepared TOP10 chemically competent cells from -80˚C freezer and thaw in ice. 2. Add between 10-100ng of DNA in solution (varies with cell competency). 3. Mix the ...)
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Revision as of 23:34, 16 June 2008
1. Remove vial of pre-prepared TOP10 chemically competent cells from -80˚C freezer and thaw in ice.
2. Add between 10-100ng of DNA in solution (varies with cell competency).
3. Mix the solution gently by tapping – anything more vigorous will result in potential damage to the competent 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. Heat shock 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 and incubate at 37˚C for one hour.
8. Plate on appropriate media and antibiotics for approximately 20 hours, then check for colonies
