Detection of specific DNA sequence through PCR

R. John Lye rjl6n at virginia.edu
Thu Sep 27 11:10:22 EST 2001


darnel wrote:

> We will be amplifying a specific DNA sequence from isolated genomic DNA
> through PCR. Our goal is to determine the presence of that gene in another
> frog specie. That specific DNA sequence is  known to be 120 bp long. If in
> our PCR experiment, we were able to amplify a fragment with nearly the same
> value (120 bp), how sure are we that we have really amplified and detected
> the same gene in another specie?? Thanks

Size alone is not sufficient to be sure that you've gotten the correct product,

especially when amplifying from another species.  You'll need to clone the
PCR product and sequence several copies and then decide whether you
have, in fact, gotten the correct fragment.  Of course, you'll need to do the
correct controls as well (no template, etc).

John







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