actin as internal standard in RT-PCR

JIrving jirving at aol.com
Thu Oct 12 02:27:33 EST 1995


Tracy;
What data are you referring to? Beside having my own empirical evidence to
go
by, I have of course tried to pay attention to the published literature on
this topic.  I would be very interested in any published work showing
general
flaws in the use of  GAPDH as an external (not internal) quantitation
standard.  I do understand that other RNA species have been proposed, but
I
have never seen ribosomal RNA listed as a candidate.  I have always
assumed
that the levels would be very sensitive to relative translational activity
of
the cell... is this incorrect?  Any good papers on this that you're aware
of?
Also, I normally wouldn't use any standard that was so highly abundant;
even
GAPDH is too abundant as a control for rare class species.  I will try to
dig
up the refs for alternative RNAs to GAPDH, I know I saw an intriguing
article
not too long ago listing several others that were claimed to be superior
(might have been in one of the in-house company publications from a
molecular
biologicals supplier).  Also, I am of course aware of the abundant GAPDH
pseudogenes, some of which are transcribed, but this does not contribute
to
variability as far as I've ever seen.  Only a handfull of papers have been
published on cataloguing the pseudogenes and sequence variants that I know
of, and none of those very recently. If you are aware of any recent papers
that do an extensive survey, I would be much obliged for the reference.

Thanks for replying,
John




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