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Title: Rapid sequencing of multiple RNA viruses in their native form
Authors: Thidathip Wongsurawat
Piroon Jenjaroenpun
Mariah K. Taylor
Jasper Lee
Aline Lavado Tolardo
Jyothi Parvathareddy
Sangam Kandel
Taylor D. Wadley
Bualan Kaewnapan
Niracha Athipanyasilp
Andrew Skidmore
Donghoon Chung
Chutikarn Chaimayo
Michael Whitt
Wannee Kantakamalakul
Ruengpung Sutthent
Navin Horthongkham
David W. Ussery
Colleen B. Jonsson
Intawat Nookaew
University of Arkansas for Medical Sciences
University of Louisville
Faculty of Medicine, Siriraj Hospital, Mahidol University
Universidade de Sao Paulo - USP
University of Tennessee Health Science Center
University of Arkansas at Little Rock
Keywords: Immunology and Microbiology
Issue Date: 1-Jan-2019
Citation: Frontiers in Microbiology. Vol.10, No.FEB (2019)
Abstract: © 2019 Frontiers Media S.A. All Rights Reserved. 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.
ISSN: 1664302X
Appears in Collections:Scopus 2019

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