miRNA-Seq of homo sapiens: adult bone marrow MSC-sEV
The total RNA of exosomes wasextracted using TRIzol (Invitrogen, Carlsbad, CA, United States).30 ng total RNA was used for library construction. TheNEBNext R Multiplex Small RNA Library kit was used toconstruct the cDNA library (NEB, Ipswich, MA, United States).The conditions of reverse transcription are set strictly accordingto the manufacturers instructions. PCR was performed underthe following cycle conditions: 94C for 30 s, 15 cycles of94C for 15 s, 62C for 30 s, and 70C for 30 s, followedby 70C for 5min. Reactions were run on an ABI Step OnePlus Real-Time PCR system. PCR products were purified usingPAGE. The constructed library was then used for quality andyield detection using an Agilent 2100 Bioanalyzer (AgilentTechnologies, Santa Clara, CA, United States). Deep sequencingwas performed on an Illumina HiSeq 2000 system. The rawdata were analyzed using FastQC1. Clean reads were annotatedin the miRBase (mature human miRNA database) to obtainthe known miRNAs.