A novel and universal method for microRNA RT-qPCR data normalization.
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.
Mestdagh
Pieter
P
Center for Medical Genetics, Ghent University Hospital, De Pintelaan 185, Ghent, Belgium. pieter.mestdagh@ugent.be
Van Vlierberghe
Pieter
P
De Weer
An
A
Muth
Daniel
D
Westermann
Frank
F
Speleman
Frank
F
Vandesompele
Jo
J
D000465:Algorithms; D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000235:genetics; D005189:False Positive Reactions; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D000071447:N-Myc Proto-Oncogene Protein; D009447:Neuroblastoma / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D018709:Statistics, Nonparametric
C459801:MYCN protein, human; D035683:MicroRNAs; D000071447:N-Myc Proto-Oncogene Protein; D009687:Nuclear Proteins; D015513:Oncogene Proteins