Brown:Transformation Protocol with our competent cells
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Revision as of 19:05, 6 July 2006
TRANSFORMATION PROTOCOL (WITH OUR COMPETENT CELLS)
1.) TAKE COMPETENT CELLS OUT OF THE -80 AND THAW (CAREFUL NOT TO THAW FOR TOO LONG)
2.) ADD 25µL CELLS TO 75µL CALCIUM CHLORIDE IN EPENDORF TUBE
3.) ADD 6µL DNA TO TUBE
4.) INCUBATE ON ICE FOR 30 MIN.
5.) HEAT SHOCK AT 42˚C FOR 1 MIN.
6.) INCUBATE ON ICE FOR 2 MIN.
7.) ADD 900µL LB/CIRCLE GROW
8.) GROW AT 37˚C WITH SHAKING FOR 1 HOUR
9.) PLATE 300µL OF MIX ONTO WARMED LB PLATES
10.) GROW OVERNIGHT AT 37˚C
