GSM6338596: CH0033; Homo sapiens; RNA-Seq
RNA extraction and clean-up from plasma was performed with Norgen Plasma/Serum RNA Purification kits (Norgen Biotek Corporation, Thorold, Canada) according to manufacturer's instructions. This kit purifies RNA from up to 5 mL of fresh or frozen serum/plasma and concentrates high purity, cell-free circulating and exosomal RNA using a two-column method. RNA was quantified using the QubitTM adapted protocol (Li et. al. BMC Mol Biol 2015, 16:9) and stored at -80°C prior to sequencing. RNA libraries were prepared using 8μl input total RNA and the SMARTer® Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Takara Bio, USA). All libraries were normalised, and equimolarly pooled before being paired-end sequenced on an Illumina NovaSeq 6000 S1 flowcell using v1.5 chemistry, generating ~100 million reads per sample at 2×100 bp read length. Library preparation was performed by the Otago Genomics Facility (Dunedin, New Zealand) and sequencing was performed by the Ramaciotti Centre for Genomics, Sydney, Australia.