title: GSM8652980: Unstim 62-A022690-L003; Homo sapiens; miRNA-Seq
library_construction_protocol: EVs obtained either from hMSCs exposed to serum-free medium or experimental samples. Evs were prepared in accordance with recent recommendations from the International Society for Extracellular Vesicles (ISEV). EV-derived miRNAs were prepared from the conditioned medium and BALF samples using the exoRNeasy Serum/Plasma Maxi/Maxi Midi Kit (Qiagen, Germantown, MD, USA), according to manufacturer's instructions. microRNA sequence was outsource to HTG and performed according to manufacture's protocols. A total of 35 ul (35 ng/µl) of eRNA (EV preparation-derived RNA) was used for miRNA sequencing performed using the HTG EdgeSeq miRNA Whole Transcriptome Assay (miRNA WTA, as per manufacturer's instructions as published). The miRNA library was sequenced on a NextSeq (Illumina, Inc., San Diego, CA) using a V3 150-cycle kit with two index reads. PhiX (Roche, Mississauga, ON, CAN). Data were returned from the sequencer in the form of demultiplexed FASTQ files, with one file per original well of the assay. The HTG EdgeSeq Parser (v. 5.0.535.3181, HTG Molecular, Tucson, AZ, USA) was used to align the FASTQ files to the probe list to collate the data. Raw read counts were imported into R to perform differential expression analyses with the R package DESeq2. The miRNA library was sequenced on a NextSeq (Illumina, Inc., San Diego, CA) using a V3 150-cycle kit with two index reads. PhiX (Roche, Mississauga, ON, CAN). Data were returned from the sequencer in the form of demultiplexed FASTQ files, with one file per original well of the assay. The HTG EdgeSeq Parser (v. 5.0.535.3181, HTG Molecular, Tucson, AZ, USA) was used to align the FASTQ files to the probe list to collate the data.