PBMC RNA-seq from a 9 month old male child, who was admitted to Kilifi county hospital in Kenya with sepsis on Tuesday, February 28, 2017 was discharged from hospital on  Thursday, March 2, 2017
Total RNA was extracted using the Qiagen RNeasy Mini Kit, according to manufacturer protocols. RNA quantity was measured using the RiboGreen assay (Invitrogen), and integrity was assessed on an Agilent 2200 TapeStation (RNA ScreenTape). RNA input was normalized to 190 ng per sample. Strand-specific mRNA libraries were prepared using the NEBNext Ultra II mRNA kit (New England Biolabs), incorporating poly-A selection. Libraries were PCR-amplified (15 cycles) with in-house unique dual indexing primers and quantified using Qubit fluorometry. Fragment size distribution was verified using the TapeStation. Normalized libraries were pooled and diluted to 10 nM for sequencing. Paired-end sequencing (2  75 bp) was performed on the Illumina HiSeq 4000 platform using HiSeq 3000/4000 PE Cluster and 150-cycle SBS kits, yielding a minimum of 19.7 million reads per sample.