Help with pcr direct from colonies

Stephen Perry bafq5 at
Fri Oct 6 08:08:40 EST 1995

I regularly use PCR to screen hundreds of transformants for inserts by
simply picking the colonies with a toothpick, streking onto a
grid-labelled plate and dipping the pick briefly into a 5ul PCR reaction
mix. I do not leave the pick and in the reaction mix as it absorbs it
really quickly!! Maybe the cause of others observed inhibition? It
always works, and makes screening very rapid. I miagine it will work
just as well for chromosomal PCR as plasmid PCR, although the lower copy
number may reqire more than the 20-25 cycles I use.

Good luck!

Steve Perry

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