GSM9453641: BMMSC-3D-EV-3; Homo sapiens; miRNA-Seq
EV samples were treated with RNase to final concentration of 50 ng/mL, at room temperature for 30 mins. RNase inhibitor and PCR grade water were added to EV samples to make a total volume of 200 μL. miRs were isolated by adding 600 μL Trizol LS according to manufacturer's instruction. To increase the yield of small RNAs, three volumes of 100% ethanol and linear acrylamide were used instead of isopropyl alcohol and incubation time was also increased to overnight at -20°C. The isolated RNAs were quantified by Qubit microRNA assay kit. Small RNA libraries (using equal RNA mass) were generated with NEBNext Multiplex Small RNA Library Prep Set for Illumina. To increase yield and prevent primer/adaptor dimer, 3' SR primer was diluted to 1:5 and ligation time was increased to overnight at 16°C.