Genome-wide identification of alternative polyadenylation events using 3′T-fill

Abstract

Due to the increasing appreciation of the impact of alternative polyadenylation on cellular biology, our straightforward, scalable method is of interest to any researcher studying eukaryotic transcription. In addition to high quality gene expression measurements, it precisely maps poly(A) sites and thereby permits the distinction between differential 3′UTR isoforms. As sequencing through long homopolymer stretches is not possible on the Illumina platform, we developed a method that fills up the poly(A) stretch with dTTPs before the sequencing reaction starts.