PCR SMEARING

malte.buchholz at medizin.uni-ulm.de malte.buchholz at medizin.uni-ulm.de
Thu Nov 9 05:41:12 EST 2000


In article <l03110703b62ff9ea8be0@[134.115.225.80]>,
  obrien at central.murdoch.edu.au ("Philip O'Brien") wrote:
> We have been getting a lot of smearing in our PCR reactions.
Sometimes we
> get a discreet band, but most of the time its a smear.  We get this
with
> different primer pairs so it dosent't appear to be confined to one
pair.
> The reactions are all set up on ice to minimize aberrant priming
before
> thermalcycling.  I would be grateful if anyone could give me
information on
> how to eliminate the problem.

What kind of PCR reaction is this, e.g. what are you trying to amplify,
what is your template material? Have you tried to vary different
reaction conditions (annealing time and temp, primer concentration etc)?
Have you tried touchdown PCR?

Cheers,

Malte

--
Dr. Malte Buchholz
malte.buchholz at medizin.uni-ulm.de


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