RiboSeq of African green monkey (Vero) cells infected at a multiplicity of infection (MOI) of 3 with the African Zika virus (ZIKV) strain, Dak84. A contamination with Toscana virus (TOSV) was discovered during bioinformatic analysis.
Vero cells were infected with the African ZIKV (Dak84) at MOI:3 to assess the viral translatome. 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. After bioinformatic analysis, it was discovered that infected cells were contaminated with another arbovirus, Toscana virus (TOSV).