Competitive PCR - Questions

douglas l feinstein dlfeins at cumc.cornell.edu
Tue Aug 23 17:51:43 EST 1994


Subject: Competitive PCR - Questions
From: Carl Waltenbaugh, cwalten at casbah.acns.nwu.edu
Date: Thu, 18 Aug 1994 18:54:55 -0500
In article <cwalten-180894185455 at 165.124.222.200> Carl Waltenbaugh,
cwalten at casbah.acns.nwu.edu writes:
>	I've been performing competitive RT-PCR on mouse DNA for a while now,
and
>have come up with a unique problem with my Interferon-Gamma analyses. 
When
>I use the primers (published by DNAX) prescribed, I get a ladder-like
>ascension of DNA bands both above AND below my expected products.  Since
>these 'ladders' get in the way of distinguishing competitor product from
RT
>product, I would like to eliminate them if possible.
>
>	Has anyone ever encountered these 'ladders' in their PCR reactions, and
if
>so, what is their origin?
>
>tg
>
>(mistert at merle.acns.nwu.edu)

Somtimes it is a hybrid formed between the true cDNA product and the
competitive standard.  Changing Mg concentration may help. Make sure your
samples are cooled , allowing all hybrids to form nicely, before running
gels.



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