Can anyone explain variable results from Ct values analysis?

05 May 2014 1 5K Report

I would like to do relative quantification of my real time PCR data, and I have designed a program based on the literature calculation and other ready programs, but the results appear variable, sometimes high or low. My sample FFPE.

Anirban Chatterjee

Dear Dhafer,

I suspect that all your FFPE samples are not uniform in their tissue type and content. What I meant to say that they might be containing variable amount of utilizable RNA. And secondly you might be be having some minute differences with your RNA isolations from your FFPE samples, which are then getting magnified in qRT-PCR.

You can go for the following Kit from Roche, it usually gives a steady result with efficient yield:

http://lifescience.roche.com/shop/products/high-pure-ffpe-rna-micro-kit#tab-1

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