RT-PCR and Template Concentration

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Thu Mar 18 04:18:10 EST 1999


In article <7cp5e9$3hv$1 at nnrp1.dejanews.com>, mbarnhart at my-dejanews.com
writes
>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.



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