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Normalization of MicroRNA Quantitative RT-PCR Data in Reduced Scale Experimental Designs

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MicroRNAs and the Immune System

Part of the book series: Methods in Molecular Biology ((MIMB,volume 667))

Abstract

Proper normalization of quantitative RT-PCR (qRT-PCR) data is a crucial component of gene ­expression analysis. Although arbitrarily selected housekeeping genes have been used to normalize many published mRNA RT-PCR datasets, there is a growing awareness that such normalizers should be first validated empirically. The use of stable reference genes is particularly needed for qRT-PCR of microRNA (miRNA), which represent a novel class of biological regulators whose aberrant expression is associated with a range of disorders. Changes in miRNA levels can be modest, and yet have profound cellular consequences. As a result, precise measurements of miRNA expression are critically important. This chapter describes a detailed workflow for the selection of endogenous normalizers using the NormFinder algorithm, ­resulting in more accurate miRNA expression profiling results. This approach is particularly well suited to smaller scale miRNA qRT-PCR experimental designs.

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Latham, G.J. (2010). Normalization of MicroRNA Quantitative RT-PCR Data in Reduced Scale Experimental Designs. In: Monticelli, S. (eds) MicroRNAs and the Immune System. Methods in Molecular Biology, vol 667. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-811-9_2

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  • DOI: https://doi.org/10.1007/978-1-60761-811-9_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-810-2

  • Online ISBN: 978-1-60761-811-9

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