title: Isolation of circulating microRNAs from microvesicles found in human plasma.
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.
authors.affiliation: Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA.
authors.forename: John F
authors.initials: JF
authors.lastname: Quackenbush
authors.forename: Pamela B
authors.initials: PB
authors.lastname: Cassidy
authors.forename: Lawrence M
authors.initials: LM
authors.lastname: Pfeffer
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Boucher
authors.forename: Jason E
authors.initials: JE
authors.lastname: Hawkes
authors.forename: Susan R
authors.initials: SR
authors.lastname: Pfeffer
authors.forename: Levy
authors.initials: L
authors.lastname: Kopelovich
authors.forename: Sancy A
authors.initials: SA
authors.lastname: Leachman
mesh_terms: D001671:Biochemistry / Q000379:methods*; D001779:Blood Coagulation Factors / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism; D002458:Cell Fractionation; D018076:DNA, Complementary / Q000096:biosynthesis; D055354:Exosomes; D006801:Humans; D016014:Linear Models; D035683:MicroRNAs / Q000097:blood* / Q000302:isolation & purification*; D010949:Plasma / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D048348:Reverse Transcription / Q000235:genetics; D022142:Secretory Vesicles / Q000378:metabolism*
chemical_list: D001779:Blood Coagulation Factors; D018076:DNA, Complementary; D035683:MicroRNAs