Multiplex Real-Time PCR Using Encoded Microparticles for MicroRNA Profiling

Seungwon Jung; Sang Kyung Kim

doi:10.1007/978-1-4939-7231-9_15

Multiplex Real-Time PCR Using Encoded Microparticles for MicroRNA Profiling

Methods Mol Biol. 2017:1654:221-230. doi: 10.1007/978-1-4939-7231-9_15.

Authors

Seungwon Jung¹, Sang Kyung Kim^{2

3}

Affiliations

¹ Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology, Seoul, South Korea.
² Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology, Seoul, South Korea. sangk@kist.re.kr.
³ Department of Biomedical Engineering, Korea University of Science and Technology, Daejeon, South Korea. sangk@kist.re.kr.

PMID: 28986793
DOI: 10.1007/978-1-4939-7231-9_15

Abstract

Multiplex quantitative real-time PCR (qPCR), which measures multiple DNAs in a given sample, has drawn unprecedented attention as a means of verifying the rapidly increasing genetic targets in a single phenotype. We report the detailed procedure of a readily extensible qPCR for multiple microRNA (miRNA) expression analysis using microparticles of primer-immobilized networks as discrete reactors. Individual particles are identified by two-dimensional codes engraved into the particles. It allows high-fidelity signal analysis in the multiplex real-time PCR. During the course of PCR, the amplicons accumulate in the volume of the particles with amplification efficiency over 95%. Tens of miRNAs can be quantitatively profiled in a single PCR reaction of this method.

Keywords: Encoded particle; Hydrogel; MicroRNA; Multiplex; Real-time PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Hydrogels / chemistry
MicroRNAs / genetics*
Real-Time Polymerase Chain Reaction / methods*

Substances

Hydrogels
MicroRNAs