Non-radioactive Quantitative RT-PCR

Harry Vlahos vlahos at mdyn.com
Thu Jan 26 20:57:38 EST 1995


shopkins at fs1.ho.man.ac.uk (Steve Hopkins) wrote:
>
> >I would like to  
> >use RT PCR to quantitate mRNA expression.  Most of the techniques I have  
> >seen require the use of radioactivity in the amplification reaction.  Are  
> >there any techniques available that a reasonably sensitive but don't  
> >require the use of radioactivity? I would appreciate any references you  
> >might have on this topic.
> 

The method of Scheuermann, R.H. and S.R. Bauer (1993) Meth. Enzymol.
218:446-473 (quantitation of oncogene messages using RT-PCR, originally
done with 32-P labeled primers) has been done successfully using both
fluorescein-labeled primers and by post-staining gels with fluorescent 
intercalating dyes such as SYBR Green.  The gels were imaged on the 
Molecular Dynamics FluorImager. (disclaimer:  Sorry to advertise--I 
work for Molecular Dynamics.)

"Siobhan Pickett" spickett at mdyn.com



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