Quantitative RT PCR method

Eric Lader lader at ambion.com
Mon Jul 28 20:10:27 EST 1997


On Thu, 24 Jul 1997 11:56:07 +0000, Andrea Itano
<aai at uclink2.berkeley.edu> wrote:

>I am trying to make my RT PCR quantitative, and I've seen one protocol 
>that recommends titering the amount of input total RNA for the RT 
>reaction, and a different protocol that recommends titering the amount 
>of cDNA template made from the RNA. Which is the more commonly used 
>method, and which would give the most quantitative comparison of the 
>relative amounts of my target mRNA?
>

What you are talking about is relative QRT-PCR. In my hands using less
RT vs using less RNA is pretty much the same. However, make every
effort to maximize sensitivity and limit cycle number. Why not give me
a call and we can chat about it. I speak faster than I type.

Eric Lader
Senior Research Scientist (the RT-PCR guy)
Ambion, Inc.
800-888-8804 x 6130



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