RT-PCR vs. RNase protection assay

Shelley Cole shelley at DARWIN.SFBR.ORG
Fri Jun 14 12:26:58 EST 1996


Several contributors to this newsgroup have stated that ribonuclease
protection assays (RPAs) are more sensitive and therefore probably
more reproducable/reliable than using quantitative RT-PCR as an assay
for gene expression.  I have been trying to convince a colleague of this,
and the usual theoretical arguments are not working. He has heard
that RT-PCR is more sensitive and a lot cheaper than RPAs. By the way,
he is a statistician/geneticist, not a wet-lab person, and he holds
the purse strings.
  Could any of you direct me to some peer-reviewed publications that
discuss comparison of these methods?  I have some review articles on
each method, but none that deal directly with comparison.
  Also, I have already developed a RPA for one of our genes of interest.
I have never done quantitative RT-PCR, so can anyone give me a realistic
estimate of the amount/time of work that would go into developing an
accurate quantitative RT-PCR assay.
  Thanks in advance.

Shelley 



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