re-amp of PCR -- problems
rafael at corona
Wed Jan 11 21:01:38 EST 1995
On 11 Jan 1995 zool201 at csc.canterbury.ac.nz wrote:
> To whom it may concern
> I'm trying to reamplify a very low concentrartion of primary PCR
> product and am ending up with smears every time.
> I don't want ot re-purify the primary product because of the very low
> Any hints would be greatfully accepted.
> Andrew Holyoake
> PhD student
> University of Canterbury
> New Zealand
Andrew, several things can try to improve your PCR
-Use different concentrations of Mg++ in the reaction (this applies for
the first reaction too). Choose the reaction with less smearing.
-Rid of the template of the first PCR, and other unwanted secondary
products: gel purification, size fractioning, etc.
-Use other internal primers to amplify in the second reaction.
I hope this helps
University of Utah
Rafael at genetics.med.utah.edu
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