optimizing RT-PCR?

Stefano casalotti stscs at mucc.mahidol.ac.th
Thu Feb 13 13:42:31 EST 1997


We have been doing quantitative RT-PCR for sometimes with reliable
results.
However one problem that we do not seem to get over is that we cannot get
1:1 realtionship between intial amount of RNA and final RT-PCR product.
We routinely check what is the suitable number of cycles for each set of
primers in order to avoid the Plateau effect. But when we do a RNA
concentration curve we see less than double RT-PCR product for every
doubling of initial RNA concentration. Some primers sets are more
problematic than others.
Has anybody figured out a trick to improve on this problem
Thanks in Advance
Stefano

Stefano Casalotti 
Neuro-behavioural Biology Center
Mahidol University, Salaya
Nakorn Pathorn 73170
Thailand
Tel  66-2)441 9321
Fax 66-2)441 9743
Email stscs at mucc.mahidol.ac.th

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