Detection of splice isoforms and rare intermediates using multiplexed primer extension sequencing. (January 2019)

• Record Type: Journal Article
• Authors: Xu, Hansen; Fair, Benjamin; Dwyer, Zachary; Gildea, Michael; Pleiss, Jeffrey
• Is Part Of: Nature methods. Volume 16:Number 1(2019)
• Page Start: 55
• Page End: 58
• Publication Date: 2019-01
• Subjects: Life sciences -- Methodology -- Periodicals; Life sciences -- Research -- Periodicals; Biology -- Methodology -- Periodicals; Biology -- Research -- Periodicals; 570.72
• DOI: 10.1038/s41592-018-0258-x
• Languages: English
• ISSNs: 1548-7091

Abstract

Abstract Targeted RNA sequencing (RNA-seq) aims to focus coverage on areas of interest that are inadequately sampled in standard RNA-seq experiments. Here we present multiplexed primer extension sequencing (MPE-seq), an approach for targeted RNA-seq that uses complex pools of reverse-transcription primers to enable sequencing enrichment at user-selected locations across the genome. We targeted hundreds to thousands of pre-mRNA splice junctions and obtained high-precision detection of splice isoforms, including rare pre-mRNA splicing intermediates. MPE-seq uses pools of reverse-transcription primers to facilitate enrichment of splice isoforms and pre-mRNA splicing intermediates.