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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