PCR problem

Chris LaRosa clarosa at biocomp.unl.edu
Mon Aug 21 09:16:49 EST 2000



Riyaz Andrabi wrote:
> 
> Hi
> I used forward and reverse primers derived from conserved sequences
> available in literature to hook out a particular gene.A single
> amplified product of 1.4kb using both the primers together as well as
> using the reverse primer alone was obtained, but no amplication was
> obtained using the forward primer alone.I want to know whether it is
> possible to hook out the full gene by using the single primer or can it
> be a part of the gene or what?Thanks
> 
Usually you need two primers.  If one alone is working the product most
likely is not the correct DNA fragment.  It is remotely possible to get
your product if there is some reverse tandem duplication , but the
chances are small.  I would stick to 2 primer strategies.






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