Increasing quality, throughput and speed of sample preparation for strand-specific messenger RNA sequencing.

ABSTRACT:

Background

RNA-Sequencing (RNA-seq) is now commonly used to reveal quantitative spatiotemporal snapshots of the transcriptome, the structures of transcripts (splice variants and fusions) and landscapes of expressed mutations. However, standard approaches for library construction typically require relatively high amounts of input RNA, are labor intensive, and are time consuming. METHODS: Here, we report the outcome of a systematic effort to optimize and streamline steps in strand-specific RNA-seq library construction. RESULTS: This work has resulted in the identification of an optimized messenger RNA isolation protocol, a potent reverse transcriptase for cDNA synthesis, and an efficient chemistry and a simplified formulation of library construction reagents. We also present an opt

SUBMITTER: Haile S 

PROVIDER: S-EPMC5499059 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature