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Normalization of Real time PCR results using Pfaffl method

PfafflReal-time PCR Serial dilution Normalization

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2 replies to this topic

#1 Benyamiin

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Posted 23 July 2012 - 10:59 PM

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

Edited by Benyamiin, 23 July 2012 - 11:08 PM.


#2 Trof

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Posted 05 August 2012 - 07:11 AM

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

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#3 thekid

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Posted 06 August 2012 - 12:51 PM

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





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