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miRNA Quantification Method Using Quantitative Polymerase Chain Reaction in Conjunction with C q Method

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Disease Gene Identification

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1706))

Abstract

MicroRNAs are small noncoding RNAs that function to regulate gene expression. In general, miRNAs are posttranscriptional regulators that imperfectly bind to the 3′untranslated region (3′UTR) of target mRNAs bearing complementary sequences, and target more than half of all protein-coding genes in the human genome. The dysregulation of miRNA expression and activity has been linked with numerous diseases, including cancer, cardiovascular diseases, neurodegenerative disorders, and diabetes. To better understand the relationship between miRNAs and human disease, a variety of techniques have been used to measure and validate miRNA expression in many cells, tissues, body fluids, and organs. For many years, quantitative polymerase chain reaction (qPCR) has been the gold standard for measuring relative gene expression, and is now also widely used to assess miRNA abundance. In this chapter, we describe a quick protocol for miRNA extraction, reverse transcription, qPCR, and data analysis.

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Acknowledgments

We thank Diego Portillo Santos for generating the figure in this chapter.

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Authors and Affiliations

  1. Center for Genes, Environment, and Health, Department of Biomedical Research, National Jewish Health, 1400 Jackson Street, Denver, CO, 80206, USA

    Fatjon Leti

  2. Translational Genomics Research Institute, 445 N 5th Street, Phoenix, AZ, 85004, USA

    Johanna K. DiStefano

Authors
  1. Fatjon Leti
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  2. Johanna K. DiStefano
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Corresponding author

Correspondence to Fatjon Leti .

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Editors and Affiliations

  1. Translational Genomics Research Institute, Phoenix, Arizona, USA

    Johanna K. DiStefano

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Leti, F., DiStefano, J.K. (2018). miRNA Quantification Method Using Quantitative Polymerase Chain Reaction in Conjunction with C q Method. In: DiStefano, J. (eds) Disease Gene Identification. Methods in Molecular Biology, vol 1706. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7471-9_14

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  • DOI: https://doi.org/10.1007/978-1-4939-7471-9_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7470-2

  • Online ISBN: 978-1-4939-7471-9

  • eBook Packages: Springer Protocols

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