Project description:Vero cells were infected with the African ZIKV (Dak84) 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. Ribosomal RNA was removed using Illumina's RiboZero kit, and RNA was hydrolysed and then 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).
Project description: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).
