Inverse PCR

Gregg Silk greggsilk at aol.com
Tue Jun 9 19:27:23 EST 1998


I successfully did inverse genomic once. You can get nothing or lots of crap,
so you need a strategy to get bands then eliminate the trash:  Cut with 4
bangers that have overhangs (Taq and several others). Try several different
enzymes. Ligate at a low DNA conc, PCR at a moderate annealling temp , using at
least 4 different primers. Try all possible primer combinations including one
at a time to identify artifact bands. Test  higher annealling temps to
eliminate the artifact bands. Takes a couple weeks, but it's straightforward.  



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