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.
journal_name
Front Microbioljournal_title
Frontiers in microbiologyauthors
Wongsurawat T,Jenjaroenpun P,Taylor MK,Lee J,Tolardo AL,Parvathareddy J,Kandel S,Wadley TD,Kaewnapan B,Athipanyasilp N,Skidmore A,Chung D,Chaimayo C,Whitt M,Kantakamalakul W,Sutthent R,Horthongkham N,Ussery DW,Jonssondoi
10.3389/fmicb.2019.00260subject
Has Abstractpub_date
2019-02-25 00:00:00pages
260issn
1664-302Xjournal_volume
10pub_type
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