GSM6043339: 9_5_3_1_CAGGCG_S1_L001; Homo sapiens; miRNA-Seq
Conditioned media samples were collected 24 h postexposure  and stored at -80 degrees C until further analysis.  Extracellular vesicles were isolated using the Invitrogen Total Exosome Isolation Reagent from Cell Culture Media (Thermo Fisher Scientific, Waltham, MA) according to manufacturer's protocol, with an additional centrifugation step at 10,000 x g for 30 minutes. RNA molecules with 18 nucleotides or greater were extracted with Invitrogen's Total Exosome RNA & Protein Isolation Kit according to manufacturer's protocol. RNA was purified and concentrated using the RNA Clean and Concentrator kit (Zymo Research, Irvine, CA) and quantified with a Nanodrop 1000 spectrophotometer (ThermoFisher Scientific, Waltham, MA) and tested for small RNA yield and quality was measured using the LabChip® Small RNA Assay (PerkinElmer). Isolated RNA samples were analyzed using next generation sequencing technologies enabled through the Illumina HiSeq 4000 platform. mRNA-seq libraries were prepared using the QuantSeq 3' mRNA-Seq Library Prep Kit for Illumina, and then these libraries were pooled and sequenced (single-end 50 bp) on the Illumina Ultra-High-Throughput Sequencing System (HiSeq) 4000.