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 sample to 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, two volume of 100% ethanol and linear acrylamide were used instead of isopropyl alcohol and incubation time was also increased to overnight at -20℃. Small RNA libraries 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 1:2 and increase ligation time to overnight at 16 ℃.
GEO Accession
GSM4697307