Quantitative RT-PCR

brett brett at BORCIM.WUSTL.EDU
Tue Oct 14 09:33:34 EST 1997

>We would like quantitate the level of gene expression by means of RT-PCR
>(Northern cannot be used since very limited amount of RNA is available).
>Anybody who is experienced in doing quantitative RT-PCR please kindly
>e-mail me the protocols. By the way, would RNase protection assay be a
>good alternative?
>Many thanks.
>Alan Lee
>e-mail: ywlee at hkucc.hku.hk

RT-PCR would be good, although you'd be wise to optimize and work out your
controls on in-vitro transcribed RNA. RPA is also good. I was able to quantitate
a viral message down to 1 copy per cell with a really hot probe.  I can supply
you with protocols if you wish.

Brett Lindenbach
Washington University - St Louis                  
brett at borcim.wustl.edu                             

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