GSM8728723: LEV, series E; Homo sapiens; RNA-Seq
Total RNA was isolated from EVs and NVEPs using Qiagen microRNA isolation kit as recommended by manufacturer. Total RNA, either 40–200 ng of EV/ NVEPs RNA, or 2 μg of cellular RNA, was rRNA-depleted using the Ribo-Zero rRNA Removal Kits (Illumina, CA). One quarter of the rRNA-depleted RNA was fragmented to 100–500 nt using the 5× First-Strand Buffer (Clontech, CA), and utilized for the RNA library construction by SMARTer Stranded RNA-Seq Kit (Clontech). The quality of libraries were examined using the Agilent DNA 1000 kit at the Agilent 2100 Bioanalyzer instrument, and cDNA quantified by qRT-PCR. The libraries were sequenced on HiSeq 2000 (Illumina).