Improving RNA-Seq expression estimates by correcting for fragment bias

Cole Trapnell
Julie Donaghey
John L Rinn
Lior Pachter
Genome Biology, 12(2010)

Abstract

The biochemistry of RNA-Seq library preparation results in cDNA fragments that are not uniformly distributed within the transcripts they represent. This non-uniformity must be accounted for when estimating expression levels, and we show how to perform the needed corrections using a likelihood based approach. We find improvements in expression estimates as measured by correlation with independently performed qRT-PCR and show that correction of bias leads to improved replicability of results across libraries and sequencing technologies.

Research Areas