Highly sensitive amplicon-based transcript quantification by semi-conductor sequencing

We have adapted a commercial assay that employs mRNA quantification based on the frequency of PCR amplicons determined by next-generation to a high-throughput semi-conductor sequencing platform (Ion-Torrent Proton). We show parallel amplification of pathway derived transcript sets/genes in 12 reference RNA samples followed by sequence-based quantification covering a dynamic range of five orders of magnitude with low technical variation. Expression of selected genes were profiled at four time points (0d, 10d, 20d, and 60d) during differentiation of induced pluripotent stem cells into cardiomyocytes, with three independent biological replicates at each time point.