title: Extensible Multiplex Real-time PCR of MicroRNA Using Microparticles.
abstract: Multiplex quantitative real-time PCR (qPCR), which measures multiple DNAs in a given sample, has received significant attention as a mean of verifying the rapidly increasing genetic targets of interest in single phenotype. Here we suggest a readily extensible qPCR for the expression analysis of multiple microRNA (miRNA) targets using microparticles of primer-immobilized networks as discrete reactors. Individual particles, 200~500 μm in diameter, are identified by two-dimensional codes engraved into the particles and the non-fluorescent encoding allows high-fidelity acquisition of signal in real-time PCR. During the course of PCR, the amplicons accumulate in the volume of the particles with high reliability and amplification efficiency over 95%. In a quick assay comprising of tens of particles holding different primers, each particle brings the independent real-time amplification curve representing the quantitative information of each target. Limited amount of sample was analyzed simultaneously in single chamber through this highly multiplexed qPCR; 10 kinds of miRNAs from purified extracellular vesicles (EVs).
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.
authors.forename: Seungwon
authors.initials: S
authors.lastname: Jung
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.|Department of Chemical &Biological Engineering, Korea University, Seoul, Korea.
authors.forename: Junsun
authors.initials: J
authors.lastname: Kim
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.
authors.forename: Dong Jin
authors.initials: DJ
authors.lastname: Lee
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.
authors.forename: Eun Hae
authors.initials: EH
authors.lastname: Oh
authors.affiliation: Center for Imaging Media Research, Robot &Media Institute, KIST, Seoul, Korea.
authors.forename: Hwasup
authors.initials: H
authors.lastname: Lim
authors.affiliation: Department of Applied Chemistry, The Institute of Natural Science, College of Applied Science, Kyung Hee University, Seoul, Korea.
authors.forename: Kwang Pyo
authors.initials: KP
authors.lastname: Kim
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.|Department of Biomedical Engineering, Korea University of Science and Technology (UST), Daejeon, Korea.
authors.forename: Nakwon
authors.initials: N
authors.lastname: Choi
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.|Department of Biomedical Engineering, Korea University of Science and Technology (UST), Daejeon, Korea.
authors.forename: Tae Song
authors.initials: TS
authors.lastname: Kim
authors.affiliation: Center for BioMicrosystems, Brain Science Institute, Korea Institute of Science and Technology (KIST), Seoul, Korea.|Department of Biomedical Engineering, Korea University of Science and Technology (UST), Daejeon, Korea.
authors.forename: Sang Kyung
authors.initials: SK
authors.lastname: Kim
mesh_terms: D055252:Cell-Derived Microparticles / Q000737:chemistry / Q000235:genetics; D000067128:Extracellular Vesicles / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D035683:MicroRNAs