Outlier detection for improved differential splicing quantification from RNA-Seq experiments with replicates

Abstract

A key component in many RNA-Seq based studies is the production of multiple replicates for varying experimental conditions. Such replicates allow to capture underlying biological variability and control for experimental ones. However, during data production researchers often lack clear definitions to what constitutes a ”bad” replicate which should be discarded and if data from failed replicates is published downstream analysis by groups using this data can be hampered. Here we develop a probability model to weigh a given RNA-Seq experiment as a representative of an experimental condition when performing alternative splicing analysis. Using both synthetic and real life data we demonstrate that this model detects outlier samples which are consistently and significantly different compared to samples from the same condition. Using both synthetic and real life data we perform extensive evaluation of the algorithm in different scenarios involving perturbed samples, mislabeled samples, no-signal groups, and different levels of coverage, and show it compares favorably with current state of the art tools.