biostudies-literatureXi SNatural Sciences and Engineering Research Council of Canada studentshipLeverhulme TrustBBSRC studentshipWellcome TrustBiotechnology and Biological Sciences Research Council222-241https://www.ebi.ac.uk/biostudies/studies/S-EPMC7618759biostudies-literatureUnknown41(4)Genomes from yeast to humans are subject to pervasive transcription. A single round of pervasive transcription is sufficient to alter local chromatin conformation, nucleosome dynamics and gene expression, but is hard to distinguish from background signals. Size fractionated native elongating transcript sequencing (sfNET-Seq) was developed to precisely map nascent transcripts independent of expression levels. RNAPII-associated nascent transcripts are fractionation into different size ranges before library construction. When anchored to the transcription start sites (TSS) of annotated genes, the combined pattern of the output metagenes gives the expected reference pattern. Bioinformatic pattern matching to the reference pattern identified 9542 transcription units in Saccharomyces cerevisiae,Yeast (Chichester, England)Size fractionated NET-Seq reveals a conserved architecture of transcription units around yeast genes.PMC7618759RPG-2016-405BB/M011224/1BB/P00296X/1WT089156MABB/S009035/1Murray SMellor JNguyen TLorenz PXi SfalseSize fractionated NET-Seq reveals a conserved architecture of transcription units around yeast genes.Genomes from yeast to humans are subject to pervasive transcription. A single round of pervasive transcription is sufficient to alter local chromatin conformation, nucleosome dynamics and gene expression, but is hard to distinguish from background signals. Size fractionated native elongating transcript sequencing (sfNET-Seq) was developed to precisely map nascent transcripts independent of expression levels. RNAPII-associated nascent transcripts are fractionation into different size ranges before library construction. When anchored to the transcription start sites (TSS) of annotated genes, the combined pattern of the output metagenes gives the expected reference pattern. Bioinformatic pattern matching to the reference pattern identified 9542 transcription units in Saccharomyces cerevisiae,2024-01-01T00:00:00Z2024 Apr2026-07-16T06:32:48.959Z2026-07-09T10:39:40.918ZS-EPMC76187593843344010.1002/yea.3931