title: whole blood-stranded RNA-Seq: Sample S16
design: RNA was extracted from blood samples (3ml) stored in Paxgene tubes using PreAnalytix kits according to manufacturers protocols. RNA (500ng) was used as input for the Ion Total RNA-Seq V2 workflow. Briefly, adapters are hybridized, and reversed transcribed. Libraries are amplified with barcoded primers (denoted in sample number) and cleaned up using AmpureXL beads. Library QC was performed using a Bioanalyzer nano DNA chip, and libraries multiplexed and seeded into a cloning PCR reaction on an OT2 (540 kit). Templated sequencing beads were enriched and loaded onto a 540 Ion Torrent sequencing chip. The chip was sequenced using the RNA-seq workflow. Each library was sequenced on two sequencing runs. Unaligned bam files from the sequencing runs were converted to fastq using picard tools (SamToFastq). For the study, data were aligned to Grch37 human reference using STAR-Bowtie2 pipeline, as part of the Ion Torrent Suite. Data were further analyzed in Partek Genomics Studio (v7).