Illumina NovaSeq 6000 paired end sequencing; Characterisation of the blood RNA host response underpinning severity in COVID-19 patients
Characterisation of the blood RNA host response underpinning severity in COVID-19 patients
Sample 12_p
Whole blood was collected at the time of recruitment into PAXgene blood miRNA tubes (PreAnalytiX), frozen, and total RNA (including RNA > 18 nucleotides) was isolated according to the manufacturer's instructions (Qiagen). RNA samples were stored at −80°C, before undergoing an additional DNAse treatment using an RNA clean & concentrator kit (Zymo Research). 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.