Quantitative RT PCR

Paul R. Rohde P.Rohde at mailbox.uq.edu.au
Thu Oct 9 21:59:01 EST 1997

lader at ambion.com writes: > In article <whc23-0710971118380001 at powercenter-604.physiol.cam.ac.uk>,
>   whc23 at cus.cam.ac.uk (Dr.W.H. Colledge) wrote:
> >
> > Can anyone please provide information or protocol for what is considered
> > the best method for quantitative RT PCR. Thanks
> In Lieu of owning a $100,000 piece of hardware...
> Assuming you are intereste in a single message: 1. design a robust pair
> of primers 2. establish a robust rt-pcr protocol providing reasonable
> yield of a single product 3. design a deletion construct and synthesize
> an RNA competitor 4. Do competitive RT-PCR with the competitor added to
> your RNA samples pre-rt.

*** Would you mind tutoring a third party ?!  What is a deletion
construct, and what is/how do you apply an RNA competitor.  I assume
you still have the same set of primers to both targets in the PCR?  ***

> well...you asked
> Eric Lader elader at ambion.com
> Senior Scientist, PCR Applications
> Ambion, Inc
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