Quick PCR

George W Chacko gchacko at magnus.acs.ohio-state.edu
Thu Apr 22 18:01:31 EST 1993


In article <9304222246.AA27633 at cscgpo> Klaus.Matthaei at anu.edu.au writes:
>Sterile toothpick colony into 100ul sterile water.
>Boil 10 min, spin eppendorf a few minutes and use 1-2ul of the supernate
>directly in your PCR reaction using your standard conditions.
>Works like a bomb.  The biggest problem is putting too much of the
>supernate into the reaction since you have too much target and the cell
>debris can inhibit the PCR.

I've barely grazed a colony with a sterile toothpick and dipped it
once or twice into the reaction mixture (through the oil) when the
thermocycler reaches 95 C. Seems to work every time.

George Chacko



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