transformation


heat shock

thaw competent cells on ice. pipet 100ul of cell suspension into a Falcon tube on ice. add plasmid DNA (.01-10ng) to cells using the pipet tip to gently mix the DNA solution and cell suspension. incubate the cells on ice for 30 min. heat shock the cells for 45 seconds in a 42C water bath. place the cells on ice for 1 min. add 900ul of LB. incubate at 37C for 1 hour, shaking at ~250 rmp. spread various aliquots of the cell suspension on LB plates containing the appropriate antibiotic. incubate at 37C overnight.


LB media

LB plates


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