Quantitative Analysis of Exosomal miRNA via qPCR and Digital PCR

Shayne A Bellingham; Mitch Shambrook; Andrew F Hill

doi:10.1007/978-1-4939-6728-5_5

Quantitative Analysis of Exosomal miRNA via qPCR and Digital PCR

Methods Mol Biol. 2017:1545:55-70. doi: 10.1007/978-1-4939-6728-5_5.

Authors

Shayne A Bellingham^{1

2}, Mitch Shambrook³, Andrew F Hill⁴

Affiliations

¹ Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, VIC, Australia.
² Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC, Australia.
³ Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Room 415, LIMS1, Bundoora, Melbourne, VIC, 3086, Australia.
⁴ Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC, Australia. Andrew.hill@latrobe.edu.au.

PMID: 27943207
DOI: 10.1007/978-1-4939-6728-5_5

Abstract

Extracellular vesicles, such as exosomes and microvesicles, have been shown to contain potential microRNA (miRNA) biomarkers that may be utilized in the diagnosis of various diseases from cancer to neurological disorders. The unique nature of the extracellular vesicle bilayer allows miRNA to be protected from degradation making it an ideal source of material for biomarkers discovery from both fresh and archived samples. Here we describe the quantitative analysis of miRNA isolated from exosomes by quantitative PCR and digital PCR.

Keywords: Biomarker; Digital PCR; Exosomes; Quantitative PCR; RNA isolation; Taqman assay; miRNA.

MeSH terms

Biomarkers
Computational Biology / methods
Exosomes / metabolism*
Gene Expression
MicroRNAs / genetics*
MicroRNAs / isolation & purification
MicroRNAs / metabolism*
RNA Transport
Real-Time Polymerase Chain Reaction*

Substances

Biomarkers
MicroRNAs