RT-PCR and Template Concentration

mbarnhart at my-dejanews.com mbarnhart at my-dejanews.com
Wed Mar 17 16:05:20 EST 1999


Hi all,

Does the amount of template RNA affect the efficiency of MMLV RT reactions?
When I have a very small amount of RNA (approx. 10-50 pg of cloned RNA
template), I get no signal in subsequent PCR reactions while I see plenty of
signal when I use a total RNA preparation containing approximately the same
number of molecules of the target RNA as in the reaction above.  The problem
might also be in the PCR reaction.  Any help would be appreciated.

Michael Barnhart
Laboratory Supervisor
Connective Tissue Physiology Lab
University of Houston

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