Nanopore native RNA sequencing of a human poly(A) transcriptome

• DOI: 10.1101/459529
• Publication Date: 2018-11-10T00:12:02Z
• Authors: Rachael E. Workman; Alison D. Tang; Paul S. Tang; Miten Jain; John R. Tyson; Philip C. Zuzarte; Timothy Gilpatrick; Roham Razaghi; Joshua Quick; Norah Sadowski; Nadine Holmes; Jaqueline Goes de...; Karen L. Jones; Terrance P. Snutch; Nicholas Loman; Benedict Paten; Matthew Loose; Jared T. Simpson; Hugh E. Olsen; Angela N. Brooks; Mark Akeson; Winston Timp
• Keywords: Nanopore

ABSTRACT High throughput cDNA sequencing technologies have dramatically advanced our understanding of transcriptome complexity and regulation. However, these methods lose information contained in biological RNA because the copied reads are often short modifications not carried forward cDNA. We address limitations using a native poly(A) strategy developed by Oxford Nanopore Technologies (ONT). Our study focused on from human cell line GM12878, generating 9.9 million aligned sequence reads. These had an N50 length 1294 bases, maximum over 21,000 bases. A total 78,199 high-confidence isoforms were identified combining long nanopore with higher accuracy Illumina describe strategies for assessing 3′ tail length, base transcript haplotypes data. Together, nanopore-based techniques poised to deliver new insights into biology. DISCLOSURES MA holds shares is paid consultant ONT. REW, WT, TG, JRT, JQ, NJL, JTS, NS, AB, MA, HEO, MJ, ML received reimbursement travel, accommodation conference fees speak at events organised NL has honorarium ONT company meeting. WT two patents (8,748,091 8,394,584) licensed Nanopore. research funding