Pfaffl calculation for qpcr: what to enter for E value? - (Jun/20/2011 )
In the calculation of relative quantification (Pfaffl 2001), the E value is replaced by E in what form? Lets say E=105%, in the formula we put E as 1.05 or 2.1? I think it should be 2.1 because if E is 100%, it'll be 1, and 1^x is always 1.
However i read an older post suggesting that it should be E=1 + amplification efficiency%; ie.: 105% > 1+1.05 = 2.05.
Can anyone verify which is the correct one? If the latter is correct, why is it so?
-hianghao-
The last suggestion seems illogical to me. I would use 2.1.
-Trof-
Trof on Tue Jun 21 10:57:08 2011 said:
The last suggestion seems illogical to me. I would use 2.1.
I thought so. I saw the latter one here:
http://www.protocol-online.org/biology-forums-2/posts/12249.html
But today i found this
"There is some ambiguity in how people define efficiency. Some people say that if you copy 90% of your DNA in a cycle, so that you end up with 1.9 times as much, the efficiency is 1.9, and this is the definition that the Pfaffl equation uses."
http://pathmicro.med.sc.edu/pcr/realtime-home.htm
-hianghao-