Chapter title |
Multiplex Real-Time PCR Using Encoded Microparticles for MicroRNA Profiling
|
---|---|
Chapter number | 15 |
Book title |
Functional Genomics
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7231-9_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7230-2, 978-1-4939-7231-9
|
Authors |
Seungwon Jung, Sang Kyung Kim |
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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