RiboSeq of African green monkey (Vero) cells infected at a multiplicity of infection (MOI) of 3 with the Asian/American Zika virus (ZIKV) strain, PE243

Vero cells were infected with the Asian/American Zika virus (ZIKV) strain, PE243, at MOI:3 to assess the viral transcriptome. Samples were harvested at 24 hours post-infection (h p.i.) by flash-freezing, without cycloheximide pre-treatment. RNase I treatment was carried out, following which ribosomes and enclosed RNA were isolated by centrifugation through a sucrose cushion. RNA was extracted, ribosomal RNA was removed using Illumina's RiboZero kit, and remaining RNA was gel purified to select fragments 25-35nt long. Fragments were cloned into adapters using the TruSeq small RNA adapter kit and sequenced on Illumina NextSeq.