title: 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.
authors.affiliation: National Center for Tumor Diseases, Heidelberg, Germany. stefan.wilkening@nct-heidelberg.de.
authors.forename: Stefan
authors.initials: S
authors.lastname: Wilkening
authors.affiliation: Genome Biology Unit, EMBL Heidelberg, Heidelberg, Germany.
authors.forename: Vicent
authors.initials: V
authors.lastname: Pelechano
authors.affiliation: Genome Biology Unit, EMBL Heidelberg, Heidelberg, Germany.|Stanford Genome Technology Center, Stanford University, Palo Alto, CA, USA.|Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Lars M
authors.initials: LM
authors.lastname: Steinmetz
mesh_terms: D020869:Gene Expression Profiling / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D011061:Poly A / Q000235:genetics*; D026723:Polyadenylation / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D059467:Transcriptome / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D011061:Poly A
keywords: Alternative polyadenylation; Dark T-fill; Expression quantification; Mapping of poly(A) sites; RNA-Seq