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Isolation of Circulating MicroRNAs from Microvesicles Found in Human Plasma

  • John F. Quackenbush
  • Pamela B. Cassidy
  • Lawrence M. Pfeffer
  • Kenneth M. Boucher
  • Jason E. Hawkes
  • Susan R. Pfeffer
  • Levy Kopelovich
  • Sancy A. Leachman
Part of the Methods in Molecular Biology book series (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.

Key words

MicroRNA Microvesicles Plasma Serum ExoQuick 

Notes

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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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • John F. Quackenbush
    • 1
  • Pamela B. Cassidy
    • 2
  • Lawrence M. Pfeffer
    • 3
  • Kenneth M. Boucher
    • 1
  • Jason E. Hawkes
    • 4
  • Susan R. Pfeffer
    • 3
  • Levy Kopelovich
    • 5
  • Sancy A. Leachman
    • 2
  1. 1.Huntsman Cancer InstituteUniversity of UtahSalt Lake CityUSA
  2. 2.Department of DermatologyOregon Health & Science UniversityPortlandUSA
  3. 3.Department of PathologyUniversity of Tennessee Health Science CenterMemphisUSA
  4. 4.Department of DermatologyUniversity of UtahSalt Lake CityUSA
  5. 5.Division of Cancer PreventionNational Cancer InstituteRockvilleUSA

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