minimum mRNA concentration for RT-PCR - (Jan/09/2008 )
hello..does anyone know what is a minimum mRNA concentration for efficient RT-PCR? tnx
-funky podmornica-
Total RNA or poly-A?
I use 1ug of total RNA. You can obviously use less but the final amount of cDNA will also decrease.
It depends on how much cDNA you'll need...
-Ambrósio-
Theoretically, one copy of a single transcript can be amplified given enough cycles in the PCR. Like ambrosio, I generally to use 1ug for RT with an oligo-dT(15) primer, though I've had success with much less input(~30ng) obtained from laser micordissected samples.
-JAH-
thanks
-funky podmornica-
We do RT-PCR on viral RNA from plasma and can get amplification when we add less than 20 copies (in case the sample was well preserved of course).
-vairus-