RT_PCR: Oligo d(T) vs. Hexamers

Pamela Norton pnorton at lac.jci.tju.edu
Wed Sep 18 13:26:41 EST 1996


In article
<c=US%a=_%p=MedImmune._Inc.%l=MEDIMMUNE_5-960916195241Z-2026 at medimmune-5.medimmune.com>,
WalshB at MEDIMMUNE.COM ("Walsh, Bill") wrote:

>         Is there an advantage to using random hexamers rather than  oligo d(T)
> when doing the first strand synthesis reaction of an RT-PCR? Is it any
> help when looking for rare transcripts?

   In a not very extensive test, I have found that random hexamers are
better than oligo dT when the PCR target sequence is a considerable
distance (a few kb) from the poly A tail. This of course makes sense.
Specific primers worked about as well, but random primers allow you to
assay multiple mRNAs from one cDNA reaction. I have no idea whether there
is a benefit when transcripts are rare.

   Pam Norton

-- 
Pamela A. Norton, Ph.D.          Assistant Professor of Medicine
Thomas Jefferson University
Philadelphia, PA 19107           p_norton at lac.jci.tju.edu



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