RT-PCR difficulty - PLEASE HELP!!!!!

Taek H. You tyou at postbox.acs.ohio-state.edu
Sat Aug 24 01:00:09 EST 1996

In article <Pine.OSF.3.93.960821121627.6082B-100000 at alpha.Colorado.EDU> Tonja Burshek <burshek at ibg.colorado.edu> writes:
>From: Tonja Burshek <burshek at ibg.colorado.edu>
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: RT-PCR difficulty - PLEASE HELP!!!!!
>Date: Wed, 21 Aug 1996 12:22:32 -0600
>Organization: University of Colorado at Boulder
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>        Background:  We are trying to reverse transcribe (following DNase
>digestion and clean-up) using an anchored poly-T primer, followed by PCR

Is this poly-T a RNA primer or you mean a poly-dT primer? 					   

>using the same poly-T paired with a 10-mer.

Can you use the same primer in PCR as in RT?
It must be a poly-dA primer to be annealed to the poly-T sequence used in cDNA 

>        Upon many moons of optimization IT WORKED BEAUTIFULLY!!!

If I am correct, that the beautiful product might turn out to be the 
ugly nonspecific (unwanted) product.

Overall, am I missing anything or misunderstanding?

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