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How to Choose a Normalization Strategy for miRNA Quantitative Real-Time (qPCR) Arrays
University of Skövde, School of Life Sciences. University of Skövde, The Systems Biology Research Centre.
University of Skövde, School of Life Sciences. University of Skövde, The Systems Biology Research Centre.
University of Skövde, School of Life Sciences. University of Skövde, The Systems Biology Research Centre.ORCID iD: 0000-0003-1837-429X
2011 (English)In: Journal of Bioinformatics and Computational Biology, ISSN 0219-7200, E-ISSN 1757-6334, Vol. 9, no 6, p. 795-812Article in journal (Refereed) Published
Abstract [en]

Low-density arrays for quantitative real-time PCR (qPCR) are increasingly being used as an experimental technique for miRNA expression profiling. As with gene expression profiling using microarrays, data from such experiments needs effective analysis methods to produce reliable and high-quality results. In the pre-processing of the data, one crucial analysis step is normalization, which aims to reduce measurement errors and technical variability among arrays that might have arisen during the execution of the experiments. However, there are currently a number of different approaches to choose among and an unsuitable applied method may induce misleading effects, which could affect the subsequent analysis steps and thereby any conclusions drawn from the results. The choice of normalization method is hence an important issue to consider. In this study we present the comparison of a number of data-driven normalization methods for TaqMan low-density arrays for qPCR and different descriptive statistical techniques that can facilitate the choice of normalization method. The performance of the normalization methods was assessed and compared against each other as well as against standard normalization using endogenous controls. The results clearly show that the data-driven methods reduce variation and represent robust alternatives to using endogenous controls.

Place, publisher, year, edition, pages
Imperial College Press, 2011. Vol. 9, no 6, p. 795-812
Keywords [en]
miRNA, qPCR array normalization, quantitative real-time PCR
National Category
Biochemistry and Molecular Biology
Research subject
Natural sciences
Identifiers
URN: urn:nbn:se:his:diva-5509DOI: 10.1142/S0219720011005793ISI: 000297096300009PubMedID: 22084014Scopus ID: 2-s2.0-80855161003OAI: oai:DiVA.org:his-5509DiVA, id: diva2:512931
Available from: 2012-03-29 Created: 2012-03-01 Last updated: 2020-05-27Bibliographically approved

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Carlsson, JessicaLindlöf, Angelica

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