GSM8879652: Control Mesenchymal stem cell-derived extracellular vesicles; Homo sapiens; miRNA-Seq
RNA was isolated from EVs using TRIzol LS reagent (10296028; Invitrogen; Thermo Fisher Scientific) according to the manufacturer's instructions. RNA quality was evaluated using an Agilent 2100 bioanalyzer equipped with an RNA 6000 Nano Chip (Agilent Technologies, Santa Clara, CA, USA). RNA quantification was performed using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific). A small RNA library was prepared using the NEBNext Multiplex Small RNA Library Prep kit (New England Biolabs, Ipswich, MA, USA) according to the manufacturer's protocol. To construct the library, cDNA synthesis was initiated by adaptor ligation using 1 μg of total RNA from each sample, followed by reverse transcription using adaptor-specific primers. The library was amplified by PCR and purified using the QIAquick PCR Purification kit (Qiagen, Hilden, Germany) and AMPure XP beads (Beckman Coulter). Then, the yield and size distribution of the small RNA libraries were analyzed using an Agilent 2100 Bioanalyzer alongside a high-sensitivity DNA assay (Agilent Technologies, USA). High-throughput sequencing was performed by single-ended 75 sequencing using the NextSeq550 system (Illumina, San Diego, CA, USA).