Poly C PCR, anyone?

Dr. Duncan Clark duncan at genesys.demon.co.uk
Sat Jul 13 04:18:42 EST 1996


In article <n1374939887.90566 at msmtp.idde.saci.org>, "\"Brett Beitzel\""
<brett_beitzel at MSMTP.FS.SACI.ORG> writes
>Subject:Poly C PCR, anyone?                        Date: 7/12/96
>
>Has anyone ever used a poly C primer for PCR (or another homopolynucleotide
>primer?)  I am trying to polyG tail a piece of DNA with Terminal transferase
>and then PCR this tailed fragment with oligo(dC)16.  I know that my TdT is
>working, but I can't get any product in my PCR.  I am wondering if there is
>something odd that happens when you try to use a homopolynucleotide primer.

It could be that because you do not know how long the G tail is (I'm
assuming it is longer than 16) that the polyC primer is priming at
multiple sites along the tail and somehow messing the PCR up. Can you
tail something that you can still PCR with internal primers? I'm just
wondering if traces of the transferase cacodylate buffer is carrying
over and inhibiting the PCR enzyme. Sorry I can't be more helpful.

Duncan 
-----------------------------------------------------------------------------
My mind's made up. Don't confuse me with the facts!
Duncan Clark
DNAmp Ltd. - Licensed PCR enzyme manufacturer
http://www.dnamp.com




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