PCR efficiencies calculation - (Jun/25/2007 )
I have to calculate PCR amplification efficiencies. Since i have many samples I will use a mix of my cDNAs and serial dilution of this mix. I am using an old ABI RT-qPCR machine and i dont have the software for data analysis. How can I calculate the slope in this case? Is there any available program for this?
Thank you very much.
Program like OpenOffice Calc of MS Excel shoud do it. (I did it once)
You just need to make a graph, where on X axis you put the common logarithm of your relative concentration and on Y axis the Ct value. The program should be able to make a regression line from the points and display the regression equation and correlation ( it should look like y= -3.7626x + 44.288 where 3.7616 is a slope I think).
Then there is the formula to calculate efficiency like 1/(log slope)? I'm not completely sure.
It is explained in ABI Prisms User Bulletin #2. Just google and you got everything you need to know.