IVT-seq reveals extreme bias in RNA sequencing.

ABSTRACT:

Background

RNA-seq is a powerful technique for identifying and quantifying transcription and splicing events, both known and novel. However, given its recent development and the proliferation of library construction methods, understanding the bias it introduces is incomplete but critical to realizing its value.

Results

We present a method, in vitro transcription sequencing (IVT-seq), for identifying and assessing the technical biases in RNA-seq library generation and sequencing at scale. We created a pool of over 1,000 in vitro transcribed RNAs from a full-length human cDNA library and sequenced them with polyA and total RNA-seq, the most common protocols. Because each cDNA is full length, and we show in vitro transcription is incredibly processive, each base in each transcr

SUBMITTER: Lahens NF 

PROVIDER: S-EPMC4197826 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature