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

Research output: Chapter in Book/Report/Conference proceedingChapter

22 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMolecular Diagnostics for Melanoma
Subtitle of host publicationMethods and Protocols
EditorsMagdalena Thurin, Francesco M. Marincola
Pages641-653
Number of pages13
DOIs
StatePublished - Jan 13 2014

Publication series

NameMethods in Molecular Biology
Volume1102
ISSN (Print)1064-3745

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Keywords

  • ExoQuick
  • MicroRNA
  • Microvesicles
  • Plasma
  • Serum

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Quackenbush, J. F., Cassidy, P. B., Pfeffer, L. M., Boucher, K. M., Hawkes, J. E., Pfeffer, S. R., Kopelovich, L., & Leachman, S. A. (2014). Isolation of circulating microRNAs from microvesicles found in human plasma. In M. Thurin, & F. M. Marincola (Eds.), Molecular Diagnostics for Melanoma: Methods and Protocols (pp. 641-653). (Methods in Molecular Biology; Vol. 1102). https://doi.org/10.1007/978-1-62703-727-3_34