PCR SMEARING

Robert Hartley rh at mblab.gla.ac.uk
Thu Nov 9 12:01:00 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.

Is this RT-PCR?

If so you could try an anchored oligo dTX or dTXX in your RT step.

Cheers
Bob; Sunny Scotland

-- 
Robert Hartley, Centre for Cell Engineering,Joseph Black Building
University of Glasgow, Glasgow G12 8QQ Tel: ++44 (0)141 330 4756, 
Fax: 0141 330 3730 mailto:rh at mblab.gla.ac.uk
Web : http://www.gla.ac.uk/Inter/CellEngineering






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