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Isolation of Circulating MicroRNAs from Microvesicles Found in Human Plasma

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Book cover Molecular Diagnostics for Melanoma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1102))

Abstract

Intact miRNAs can be isolated from the circulation in significant quantities despite the presence of extremely high levels of RNase activity. The remarkable stability of circulating miRNAs makes them excellent candidates for biomarkers in diagnostic applications as well as therapeutic targets in a variety of disease states including melanoma. Circulating RNA molecules are resistant to degradation by RNases because they are encapsulated in membrane-bound microvesicles. We describe a convenient method for the use of ExoQuick, a proprietary resin developed by Systems Biosciences (Mountain View, CA), whereby microvesicles can be purified under gentle conditions using readily available laboratory equipment. This protocol allows for isolation all microvesicles, regardless of their origin, and provides a convenient method for identifying potential cancer-specific biomarkers from biological fluids including serum and plasma.

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Acknowledgment

This work was supported by a subcontract with the NCI number N201143, the Tom C. Mathews Familial Melanoma Research Clinic, the Huntsman Cancer Foundation, and the Cancer Center Support Grant for the University of Utah 5P30CA042014-23.

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Authors and Affiliations

  1. Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA

    John F. Quackenbush & Kenneth M. Boucher

  2. Department of Dermatology, Oregon Health & Science University, Portland, OR, USA

    Pamela B. Cassidy & Sancy A. Leachman

  3. Department of Pathology, University of Tennessee Health Science Center, Memphis, TN, USA

    Lawrence M. Pfeffer & Susan R. Pfeffer

  4. Department of Dermatology, University of Utah, Salt Lake City, UT, USA

    Jason E. Hawkes

  5. Division of Cancer Prevention, National Cancer Institute, Rockville, MD, USA

    Levy Kopelovich

Authors
  1. John F. Quackenbush
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  2. Pamela B. Cassidy
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  3. Lawrence M. Pfeffer
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  4. Kenneth M. Boucher
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  5. Jason E. Hawkes
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  6. Susan R. Pfeffer
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  7. Levy Kopelovich
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  8. Sancy A. Leachman
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Editor information

Editors and Affiliations

  1. Cancer Diagnosis Program, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    Magdalena Thurin

  2. Sidra Medical and Research Center,Qatar Foundation, Doha, Qatar

    Francesco M. Marincola

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© 2014 Springer Science+Business Media, New York

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Quackenbush, J.F. et al. (2014). Isolation of Circulating MicroRNAs from Microvesicles Found in Human Plasma. In: Thurin, M., Marincola, F. (eds) Molecular Diagnostics for Melanoma. Methods in Molecular Biology, vol 1102. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-727-3_34

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  • DOI: https://doi.org/10.1007/978-1-62703-727-3_34

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-726-6

  • Online ISBN: 978-1-62703-727-3

  • eBook Packages: Springer Protocols

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