Whole-blood RNA-seq of adults with sepsis in Uganda
From whole-blood samples collected in PAXgene RNA tubes (PreAnalytiX, Qiagen / BD), RNA was isolated and purified using PAXgene blood RNA kits (Qiagen). DNase treatment was performed using Turbo DNase (Thermo Fisher Scientific) prior to library preparation. rRNA depletion was performed using QIAseq FastSelect  rRNA/Globin Kit (Qiagen). RNA sequencing library preparation was performed using the NEBNext Ultra RNA Library Prep Kit for Illumina following the manufacturers recommendations (NEB). Sequencing libraries were validated using the 4200 Tapestation (Agilent Technologies) and quantified using the Qubit 2.0 Fluorometer (Invitrogen, Thermo Fisher Scientific) as well as by quantitative PCR (Applied Biosystems, Thermo Fisher Scientific). Sequencing libraries were multiplexed and clustered on one lane of a flowcell and loaded on the Illumina HiSeq instrument (Illumina, Inc.) according to manufacturers instructions. Samples were sequenced using a 2x150 paired end configuration.