Isolation of circulating microRNAs from microvesicles found in human plasma

John F. Quackenbush, Pamela B. Cassidy, Lawrence Pfeffer, Kenneth M. Boucher, Jason E. Hawkes, Susan R. Pfeffer, Levy Kopelovich, Sancy A. Leachman

Research output: Chapter in Book/Report/Conference proceedingChapter

20 Citations (Scopus)

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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Ribonucleases
MicroRNAs
Tumor Biomarkers
Melanoma
Biomarkers
RNA
Equipment and Supplies
Membranes
Serum
Therapeutics

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Quackenbush, J. F., Cassidy, P. B., Pfeffer, L., Boucher, K. M., Hawkes, J. E., Pfeffer, S. R., ... 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

Isolation of circulating microRNAs from microvesicles found in human plasma. / Quackenbush, John F.; Cassidy, Pamela B.; Pfeffer, Lawrence; Boucher, Kenneth M.; Hawkes, Jason E.; Pfeffer, Susan R.; Kopelovich, Levy; Leachman, Sancy A.

Molecular Diagnostics for Melanoma: Methods and Protocols. ed. / Magdalena Thurin; Francesco M. Marincola. 2014. p. 641-653 (Methods in Molecular Biology; Vol. 1102).

Research output: Chapter in Book/Report/Conference proceedingChapter

Quackenbush, JF, Cassidy, PB, Pfeffer, L, Boucher, KM, Hawkes, JE, Pfeffer, SR, Kopelovich, L & Leachman, SA 2014, Isolation of circulating microRNAs from microvesicles found in human plasma. in M Thurin & FM Marincola (eds), Molecular Diagnostics for Melanoma: Methods and Protocols. Methods in Molecular Biology, vol. 1102, pp. 641-653. https://doi.org/10.1007/978-1-62703-727-3_34
Quackenbush JF, Cassidy PB, Pfeffer L, Boucher KM, Hawkes JE, Pfeffer SR et al. Isolation of circulating microRNAs from microvesicles found in human plasma. In Thurin M, Marincola FM, editors, Molecular Diagnostics for Melanoma: Methods and Protocols. 2014. p. 641-653. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-727-3_34
Quackenbush, John F. ; Cassidy, Pamela B. ; Pfeffer, Lawrence ; Boucher, Kenneth M. ; Hawkes, Jason E. ; Pfeffer, Susan R. ; Kopelovich, Levy ; Leachman, Sancy A. / Isolation of circulating microRNAs from microvesicles found in human plasma. Molecular Diagnostics for Melanoma: Methods and Protocols. editor / Magdalena Thurin ; Francesco M. Marincola. 2014. pp. 641-653 (Methods in Molecular Biology).
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