Good HKG for quantitative PCR

Andre Hamel hamel at ccu.umanitoba.ca
Tue Feb 23 14:52:23 EST 1993


>know of a gene or genes (of house keeping variety) other than actin or
>G3PD that can be used for standardizing quantitative PCR from
>RTed mRNA extracted from mammalian tissue samples?  The problem
>with these two genes, I gather, are the psuedo genes that amp
>from the DNA also found the nucleic acid prep. There has been no luck 
>so far with with DNAseI digestion, followed by phenol extraction and 
>precipitation (using glycogen as a carrier).  A simple fix is to use a
house keeping gene that has no psuedo genes.  Are there any currently used
for this purpose? Thanks in advance!!!!

Would 28S ribsomal RNA gene be suitable? Seem to recall several papers
referring to rRNA as standardizing background marker for quantitating
mRNA. One could run RT PCR on total RNA before and after mRNA purification
to confirm this (and/or use non-polyA RNA (unbound "flow-thru") after mRNA
purification.

Andre



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