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Normalization of Real time PCR results using Pfaffl method - (Jul/24/2012 )

Greetings every one,

I am planning to do a Real-Time PCR, and I thought of using Pfaffl method to normalize my results, Since the amount of samples I can get from my subjects are very limited, I can not use them to prepare serial dilution and get the standard curve to calculate the efficiency.
I wonder if I can pool samples of some of my controls, and get the standard curve using that.

Cheers and thank you


You don't need samples to do a dilution curve. Any template similar to your sample would do. (or pool, yes)



AM planning to do expression studies too and would like to use the pfaffl method for data analysis. But I am doing my experiments on a ABI 7500 machine which calculates based on the delta delta ct method. Woul like to know if I can calculate the pfaffl method using the raw data. I did come across a program called Qbase plus but it is a paid software. Would like to know if there is a free software that can be used to calculate the pfaffl method using the raw data.