title: Raw unaligned whole-blood RNAseq data
design: Total RNA was extracted from whole blood samples in PAXgene blood RNA tubes using PAXgene Blood RNA Kits (cat. no. 762164) (Qiagen, Germantown, MD, USA) in accordance with the manufacturer's protocol (Azenta Life Sciences, South Plainfield, NJ, USA). Total RNA samples were quantified using a Qubit Fluorometer 3.0 (Life Technologies, Carlsbad, CA, USA; ThermoFisher Scientific, Waltham, MA, USA). RNA integrity was checked with a 4200 TapeStation (Agilent Technologies, Palo Alto, CA, USA). Samples were initially treated with TURBO DNase (cat. no. AM2238) (Thermo Fisher Scientific, Waltham, MA, USA) to remove DNA contaminants, after which rRNA and globin depletion were performed using QIAseq FastSelect−rRNA HMR and −Globin kits (cat. no. 334375) (Qiagen). RNA sequencing libraries were constructed with the NEBNext Ultra II RNA Library Preparation Kit for Illumina by following the manufacturer's recommendations. Sequencing libraries were validated using the Agilent Tapestation 4200 and quantified using a Qubit Fluorometer 3.0 and quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). Sequencing libraries were multiplexed and clustered onto a flowcell on the Illumina NovaSeq X Plus instrument according to manufacturer's instructions. Samples were sequenced using a 2x150bp paired end configuration. Image analysis and base calling were conducted by NovaSeq Control. Raw sequence data (bcl files) was converted into fastq files and de-multiplexed using Illumina bcl2fastq software 2.20
identifiers.namespace: NIAID-phs003914
identifiers.id: CL-601-TRO