Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR.
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Marabita F, de Candia P, Torri A, Tegnér J, Abrignani S, Rossi RL
Brief. Bioinformatics 17 (2) 204-212 [2016-03-00; online 2015-08-03]
The high-throughput analysis of microRNAs (miRNAs) circulating within the blood of healthy and diseased individuals is an active area of biomarker research. Whereas quantitative real-time reverse transcription polymerase chain reaction (qPCR)-based methods are widely used, it is yet unresolved how the data should be normalized. Here, we show that a combination of different algorithms results in the identification of candidate reference miRNAs that can be exploited as normalizers, in both discovery and validation phases. Using the methodology considered here, we identify normalizers that are able to reduce nonbiological variation in the data and we present several case studies, to illustrate the relevance in the context of physiological or pathological scenarios. In conclusion, the discovery of stable reference miRNAs from high-throughput studies allows appropriate normalization of focused qPCR assays.
PubMed 26238539
DOI 10.1093/bib/bbv056
Crossref 10.1093/bib/bbv056
pii: bbv056
pmc: PMC4793896