If you start off with 20ul of 20ng/ul of total RNA, and you need 4ul of this for a 20ul cDNA reaction (4ul template RNA + 16ul of enzyme and buffer mix).
My understanding is I've input 4ul of 20ng/ul total RNA, so that's 80ng of 20ng/ul.
How much cDNA will my output amount be?
David Roig-Carles
Hey,
cDNA reaction is 1:1 ratio...therefore you get 80ng of cDNA. It doesn't amplify your RNA. You can always double check your cDNA concentration using nanodrop afterwards.
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Alberto Angrisani
Hi Marc,
just a clarification, the 1:1 ratio is correct, but only theoretical for random hexamers primed reaction.
If you prime cDNA syntesis with oligo-dT your yeld will be much less, in theory only mRNAs, about 2% of the total RNA.
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