Rapid Sequencing of Multiple RNA Viruses in their Native Form
Mariah K Taylor,
Aline Lavado Tolardo,
Taylor D Wadley,
David W. Ussery,
Colleen B Jonsson,
Posted 29 Nov 2018
bioRxiv DOI: 10.1101/482471 (published DOI: 10.3389/fmicb.2019.00260)
Posted 29 Nov 2018
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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