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Reference gene(s) selection and validaton for qPCR

housekeeping normalization reference

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#1 Trof


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Posted 22 April 2014 - 05:01 AM

Selecting a proper reference gene(s) for RT-qPCR is an ongoing struggle.

It's easy to "do it", but it's significantly more difficult to do it right.


I will dedicate this topic to general discussion about selecting reference (sometimes called housekeeping) genes, software/application used and so on.


A web page first.


Massively resourced but rather ugly pages concerning normalization and reference genes selection on Gene-Quantification.info site.



(often contains fulltexts of papers)


Some papers.


The need for transparency and good practices in the qPCR literature - Nature Methods 2013

Evidence Based Selection of Housekeeping Genes - PloS ONE 2007


and of course


MIQE Guidelines


Today I came accross a free web tool to select reference genes using algorithms from four currently used applications geNorm, Normfinder, BestKeeper, and the comparative ddCt method.

Since I have used only one of them and this requires to only copy Ct directly from the Excel table (!!) it's like a huge help to refgene selection (I used GeNorm for some time, I wouldn't really call it user friendly..)



But since I can't compare with the others I don't have any comments on this would be appreciated.


(I may add later some sites/papers/... I found highly relevant, post any sources regarding the topic, if you want, too)

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

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#2 Inbox



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Posted 18 July 2014 - 07:39 AM

Hey Trof, 

       Thanks for quiet useful information. Could you please add material you mentioned in last post?

Edited by Inbox, 18 July 2014 - 07:39 AM.

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