Two-pass alignment improves novel splice junction quantification.

ABSTRACT:

Motivation

Discovery of novel splicing from RNA sequence data remains a critical and exciting focus of transcriptomics, but reduced alignment power impedes expression quantification of novel splice junctions.

Results

Here, we profile performance characteristics of two-pass alignment, which separates splice junction discovery from quantification. Per sample, across a variety of transcriptome sequencing datasets, two-pass alignment improved quantification of at least 94% of simulated novel splice junctions, and provided as much as 1.7-fold deeper median read depth over those splice junctions. We further demonstrate that two-pass alignment works by increasing alignment of reads to splice junctions by short lengths, and that potential alignment errors are readily identifiable by

SUBMITTER: Veeneman BA 

PROVIDER: S-EPMC5006238 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature