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.
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Acknowledgments
This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (Grant No. HI13C2262). This work was also supported by KIST through the Institutional Program (Project No. 2E25590) and Open Research Program (Project No. 2E25722).
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Jung, S., Kim, S.K. (2017). Multiplex Real-Time PCR Using Encoded Microparticles for MicroRNA Profiling. In: Kaufmann, M., Klinger, C., Savelsbergh, A. (eds) Functional Genomics. Methods in Molecular Biology, vol 1654. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7231-9_15
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DOI: https://doi.org/10.1007/978-1-4939-7231-9_15
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