Targeted RNA-sequencing with competitive multiplex-PCR amplicon libraries.

ABSTRACT: Whole transcriptome RNA-sequencing is a powerful tool, but is costly and yields complex data sets that limit its utility in molecular diagnostic testing. A targeted quantitative RNA-sequencing method that is reproducible and reduces the number of sequencing reads required to measure transcripts over the full range of expression would be better suited to diagnostic testing. Toward this goal, we developed a competitive multiplex PCR-based amplicon sequencing library preparation method that a) targets only the sequences of interest and b) controls for inter-target variation in PCR amplification during library preparation by measuring each transcript native template relative to a known number of synthetic competitive template internal standard copies. To determine the utility of this method, w

SUBMITTER: Blomquist TM 

PROVIDER: S-EPMC3827295 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature