Rapid Sequencing of Multiple RNA Viruses in Their Native Form

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, subge...

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Detalles Bibliográficos
Autores principales: Wongsurawat, Thidathip, Jenjaroenpun, Piroon, Taylor, Mariah K., Lee, Jasper, Tolardo, Aline Lavado, Parvathareddy, Jyothi, Kandel, Sangam, Wadley, Taylor D., Kaewnapan, Bualan, Athipanyasilp, Niracha, Skidmore, Andrew, Chung, Donghoon, Chaimayo, Chutikarn, Whitt, Michael, Kantakamalakul, Wannee, Sutthent, Ruengpung, Horthongkham, Navin, Ussery, David W., Jonsson, Colleen B., Nookaew, Intawat
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Microbiology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6398364/
https://www.ncbi.nlm.nih.gov/pubmed/30858830
http://dx.doi.org/10.3389/fmicb.2019.00260
Descripción
Sumario: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.

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