Genome-Seq of DENV-2:Vanuatu
The extracted viral RNA was circularized to enable complete sequencing of the terminal 5 and 3 untranslated regions using tobacco decapping enzyme (Enzymax LLC, USA) and T4 RNA ligase (New England Biolabs, Genesearch, Arundel, Australia) as reported previously. To remove host and potentially contaminating microbial DNA, the circularized RNA was then DNase treated (Heat & Run Kit, ArcticZymes, Scientifix, South Yarra, Australia). WGS (Illumina, San Diego, CA) was performed as described previously using the Nextera XT kit for cDNA library construction followed by paired-end (2  151 nt) sequencing using the V2 mid-output kit on a NextSeq 500 machine.