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Normalization of Real time PCR results using Pfaffl method - (Jul/23/2012 )

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

-Benyamiin-

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

-Trof-

Hello,

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.


regards,
thekid

-thekid-