title: RNA-Seq of healthy adult volunteers following experimental infection with Influenza A/Belgium/4217/2015 (H3N2)
design: RNA-Seq of healthy adult volunteers following experimental infection with Influenza A/Belgium/4217/2015 (H3N2)
library_construction_protocol: Nasal curettage samples were collected at baseline, Days 1, 2, 3, 7, 10 and 14 post-inoculation and preserved in TRIzol. These were stored at -80°C until processing. Total RNA was extracted from nasal curettage samples using Qiagen RNeasy Mini kit (Qiagen, Hilden, Germany). Strand-specific RNA sequencing library was prepared by using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB, Ipswich, MA, USA). The enriched RNAs were fragmented for 8 minutes at 94 °C. The first strand and second strand cDNA were subsequently synthesized. The second strand of cDNA was marked by incorporating dUTP during the synthesis. cDNA fragments were adenylated at 3'ends, and indexed adapter was ligated to cDNA fragments. Limited cycle PCR was used for library enrichment.