Internal RT-PCR controls

Dr. Duncan Clark drc at nospam.demon.co.uk
Thu Jun 29 10:46:39 EST 2000


In article <395B2FFF.DDF600F8 at sheffield.ac.uk>, Lee Hunt
<L.hunt at sheffield.ac.uk> writes
>Can I use my internal control primers in the same PCR reaction as my
>specific primers, 

Yes solely providing the primer pairs do not interact. 

>and is this more quantitiative than doing the 2
>reactions separately?

Probably, but only if the primer pairs amplify their targets with the
same efficiencies and that one primer pair does not out compete the
second pair for reagents. That would mean that the product bands for
each target should appear at similar cycle numbers. A bit of work will
need doing to check this out before you rely on the result.

Duncan 

-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk






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