Extensible Multiplex Real-time PCR of MicroRNA Using Microparticles
- Extensible Multiplex Real-time PCR of MicroRNA Using Microparticles
- 정승원; 김준선; 이동진; 오은해; 임화섭; 김광표; 최낙원; 김태송; 김상경
- 분자진단; 다중마커 진단; 실시간 PCR; 입자기반 qPCR; miRNA; 패턴인식 입자식별
- Issue Date
- Scientific Reports
- VOL 6, NO 22975
- 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).
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