Multiplex Real-Time PCR Using Encoded Microparticles for MicroRNA Profiling.

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.