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A novel and universal method for microRNA RT-qPCR data normalization

Pieter Mestdagh1, Pieter Van Vlierberghe1, An De Weer1, Daniel Muth2, Frank Westermann2, Frank Speleman1 and Jo Vandesompele1*

Author Affiliations

1 Center for Medical Genetics, Ghent University Hospital, De Pintelaan 185, Ghent, Belgium

2 Department of Tumour Genetics, German Cancer Center, Im Neuenheimer Feld 280, Heidelberg, Germany

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Genome Biology 2009, 10:R64  doi:10.1186/gb-2009-10-6-r64

Published: 16 June 2009

Abstract

Gene expression analysis of microRNA molecules is becoming increasingly important. In this study we assess the use of the mean expression value of all expressed microRNAs in a given sample as a normalization factor for microRNA real-time quantitative PCR data and compare its performance to the currently adopted approach. We demonstrate that the mean expression value outperforms the current normalization strategy in terms of better reduction of technical variation and more accurate appreciation of biological changes.