| Literature DB >> 30858830 |
Thidathip Wongsurawat1, Piroon Jenjaroenpun1, Mariah K Taylor2, Jasper Lee2, Aline Lavado Tolardo3, Jyothi Parvathareddy4, Sangam Kandel1,5, Taylor D Wadley1, Bualan Kaewnapan6, Niracha Athipanyasilp6, Andrew Skidmore7, Donghoon Chung7, Chutikarn Chaimayo6, Michael Whitt2, Wannee Kantakamalakul6, Ruengpung Sutthent6, Navin Horthongkham6, David W Ussery1,8, Colleen B Jonsson2, Intawat Nookaew1,8.
Abstract
Long-read nanopore sequencing by a MinION device offers the unique possibility to directly sequence native RNA. We combined an enzymatic poly-A tailing reaction with the native RNA sequencing to (i) sequence complex population of single-stranded (ss)RNA viruses in parallel, (ii) detect genome, subgenomic mRNA/mRNA simultaneously, (iii) detect a complex transcriptomic architecture without the need for assembly, (iv) enable real-time detection. Using this protocol, positive-ssRNA, negative-ssRNA, with/without a poly(A)-tail, segmented/non-segmented genomes were mixed and sequenced in parallel. Mapping of the generated sequences on the reference genomes showed 100% length recovery with up to 97% identity. This work provides a proof of principle and the validity of this strategy, opening up a wide range of applications to study RNA viruses.Entities:
Keywords: MinION; genome; nanopore sequencing; native RNA; rapid detection; single-stranded RNA; subgenomic mRNA; virus
Year: 2019 PMID： 30858830 PMCID： PMC6398364 DOI： 10.3389/fmicb.2019.00260
Source DB: PubMed Journal: Front Microbiol ISSN： 1664-302X Impact factor: 5.640