Whole Blood RNAseq of RAS Vaccine Cohort

Whole blood RNAseq was performed as previously described (13 (link)) on 11 IMRAS Cohort 1 RAS vaccinees. Whole blood was collected directly into PAXgene blood RNA tubes (PreAnalytiX, Hombrechtikon, Switzerland) and stored at −20°C. RNA extraction and globin transcript depletion (GlobinClear, ThermoFisher Scientific, MA, USA) were performed prior to cDNA library preparation using the Illumina TruSeq Stranded mRNA sample preparation kit (Illumina, CA, USA). Globin transcript depletion, cDNA library preparation and RNA sequencing were performed by the Beijing Genomics Institute (Shenzhen, China). A total of sixty-six RNAseq samples were sequenced, with a target depth of 30 million reads per sample. Samples from four timepoints (day 0, 35, 119, 140) were sequenced on Illumina (San Diego, CA) Hiseq2000 sequencers using 75 base-pair (bp) paired-end reads. The remaining samples were sequenced on BGI500 sequencers using 100 bp paired-end reads.