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

Taek H. You tyou at postbox.acs.ohio-state.edu
Tue Aug 27 23:31:33 EST 1996

In article <tyou.320.321E9A69 at postbox.acs.ohio-state.edu> tyou at postbox.acs.ohio-state.edu (Taek H. You) writes:
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>From: tyou at postbox.acs.ohio-state.edu (Taek H. You)
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: Re: RT-PCR difficulty - PLEASE HELP!!!!!
>Date: Sat, 24 Aug 1996 06:00:09 GMT
>Organization: The Ohio State University
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>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?

I've made a mistake on this.
I only thought about PCR, not considering RT reaction.
Did you used a specific primer in PCR?

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