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microRNAs and Reference Gene Methodology

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Handbook of Nutrition, Diet, and Epigenetics

Abstract

microRNAs (miRNAs), short noncoding RNAs, are posttranscriptional negative regulators, with extracellular circulating miRNAs considered promising biomarkers of various diseases. The relative quantification of miRNA transcripts requires an endogenous reference gene for data normalization, which is the critical step in this process. In the present chapter, the means of normalization and methods for the selection of suitable reference gene(s) are discussed, with a key finding being how common reference genes such as RNU6 and miR-16 can cause bias in data interpretation if used without proper validation.

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Abbreviations

18S:

Ribosomal RNA 18S

5S:

Ribosomal RNA 5S

ACT:

Actin

Cq:

Quantitation cycle

FFPE:

Formalin-fixed paraffin-embedded

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

LNA:

Locked nucleic acid

MIQE:

Minimum information for publication of quantitative real-time PCR experiments

miRNA:

microRNA

qPCR:

Quantitative real-time polymerase chain reaction

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Authors and Affiliations

  1. Department of Biochemical Sciences, Charles University, Faculty of Pharmacy, Akademika Heyrovského 1203/8, 50005, Hradec Králové, Czech Republic

    Petra Matoušková

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  1. Petra Matoušková
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Correspondence to Petra Matoušková .

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Editors and Affiliations

  1. c/o Editorial Board 978-0-387-78665 , Heidelberg, Baden-Württemberg, Germany

    Vinood Patel

  2. Kings College , London, United Kingdom

    Victor Preedy

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Matoušková, P. (2017). microRNAs and Reference Gene Methodology. In: Patel, V., Preedy, V. (eds) Handbook of Nutrition, Diet, and Epigenetics. Springer, Cham. https://doi.org/10.1007/978-3-319-31143-2_34-1

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  • DOI: https://doi.org/10.1007/978-3-319-31143-2_34-1

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