title: Illumina NovaSeq 6000 paired end sequencing; Analysis of blood and nasal epithelial transcriptomes to identify mechanisms associated with control of SARS-CoV-2 viral load in the upper respiratory tract
design: Analysis of blood and nasal epithelial transcriptomes to identify mechanisms associated with control of SARS-CoV-2 viral load in the upper respiratory tract
library_construction_protocol: Whole blood was collected at the time of recruitment into PAXgene blood miRNA tubes (PreAnalytiX) and frozen. Total RNA (including RNA > 18 nucleotides) was isolated using RNeasy microkit according to the manufacturer's instructions (Qiagen). Input material was normalised and strand specific library preparation was completed using NEBNext Ultra II mRNA kit (NEB) and NEB rRNA/globin depletion probes following manufacturer's instructions. Libraries were on a Tetrad (Bio-Rad) using in-house unique dual indexing primers. Individual libraries were normalised using Qubit and pooled together. The pooled library was diluted to ~10 nM for storage and denatured and further diluted prior to loading on the sequencer.