GSM7874154: control EVs #1; Homo sapiens; RNA-Seq
Total RNA was extracted from the isolated ITGA6+EVs and control EVs using Trizol (Invitrogen, Carlsbad, CA, USA) according to the manual instructions. Total RNA was qualified and quantified using a Nano Drop and Agilent 2100 bioanalyzer (Thermo Fisher Scientific, MA, USA). Then, 3μg of total RNA was treated with DNase I which degraded double-stranded and single-stranded DNA presenting in RNA samples. And the ribosomal RNA was removed using the Ribo-off rRNA Depletion Kit (Vazyme, Inc.). Linear RNA was removed using RNase R (Epicentre, lnc). Purification was performed using Agencourt RNAClean XP magnetic beads. All other steps were performed according to the manufacturer's protocols. The library was quality and quantitated in two methods: check the distribution of the fragments size using the Agilent 2100 bioanalyzer, and quantify the library using BMG (OMEGA). RNA libraries were prepared for sequencing using standard DNBSEQ protocols