GSM4257615: Fish, EVs,  3 Day 1; Salmo salar; miRNA-Seq
Extracellular vesicles were isolated from the culture media using VN96 (New England Peptide) following the manufacturer's protocol. Total RNA was extracted using the mirVana kit (Invitrogen) following the manufacturer's protocol. RNA quality was assessed on Fragment analyzer using HS RNA assay and HS small RNA assay (Agilent Technologies). Small RNA library was prepared using Clean Tag Small RNA library prep kit following manufacturer recommended conditons (TriLink Biotech). Ion Torrent specific RT primer and barcodes were used during the library construction. Barcoded cDNA libraries were subjected to double size selection using Ampure XP beads (Beckman Coulter) to enrich for miRNA transcripts.  Quality of the library was analyzed using D1000 assay on Tapestation 2200 (Agilent Technologies). Libraries were equally pooled at a loading concentration of 7pM and amplified onto Ion Sphere™ Particles (Ion Pi HiQ OT2 kit, Life Technologies). ISP enriched library was sequenced using Ion Proton (ThermoFisher).
GEO
GSM4257615