Good HKG for quantitative PCR

Jorge L. Sepulveda Jorges at bcm.tmc.edu
Mon Feb 22 17:58:09 EST 1993


In article <2B870CD4 at adminbldg.lan1.umanitoba.ca> 
GIETZ at bldghsc.lan1.umanitoba.ca writes:
> Hello Fellow Netters,
>     I am posting this question for a friend
> who is not hard wired to the net yet.  Does anyone
> 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?

If you pick up primers from different exons (i.e.) separated by introns in 
the genomic DNA, there should be no problem with unexpressed genes.

Cheers,


Jorge L. Sepulveda, 

Dept. of Pathology,                        Email: Jorges at bcm.tmc.edu
Baylor College of Medicine                 Telephone: 713-798-7330
One Baylor Plaza, Houston, TX 77030        Telefax: 713-798-5838



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