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How to Normalize of RT-qPCR with several reference genes ?


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#1 Meirp

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Posted 04 March 2010 - 07:27 AM

Hi,
I've performed a RT-qPCR analysis with one target gene and two reference genes. Now I would like to calculate the mean normalized expression (MNE') of my target gene (Muller et al. 2002) using a normalization by multiple HKG (Vandesompele et al. 2002). My question is what should I do to adjust the calculation with the PCR efficiencies of my references? Should I take the geometric mean of the PCR efficiencies of my two reference genes?

Thanks in advance for your help!

#2 tea-test

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Posted 04 March 2010 - 09:23 AM

Hi,

if you want to normalize to more than one reference gene and adjust for different efficiences you need a different approach. you can find the formulas here:

pubmed link
tea-test: The artist formerly known as Ned Land

#3 Meirp

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Posted 05 March 2010 - 01:36 AM

Thank you for your quick answer, it was really helpful !




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