GSM8843761: Human FF-EVs, Aged, Rep 3; Homo sapiens; miRNA-Seq
Follicular fluid EV samples were thawed on ice and total RNA was isolated using an Exosomal RNA Isolation Kit (Norgen Biotek Corp., Cat. 58000) according to the manufacturer's instructions. The RNA integrity and concentration were measured using an Agilent 2100 Bioanalyzer (Agilent Technologies, CA) and a NanoDrop™ 2000 Spectrophotometer (ThermoFisher Scientific, Cat. ND-2000), respectively. The isolated total RNA was sent to Novogene Co. LTD (California, USA) for library preparation and sequencing. The small-RNA libraries were constructed using the TruSeq Small RNA Library Prep Kit (Illumina RS-200) according to the manufacturer's instructions. Library quantity and quality measurements were taken using a Qubit® DNA HS Assay Kit on a Qubit® 2.0 Fluorometer and an Agilent DNA High Sensitivity Kit in an Agilent 2100 Bioanalyzer, respectively. Library concentrations were calculated using quantitative PCR, then pooled in equimolar ratios, and sequenced on a NovaSeq6000 sequencing instrument (Illumina) as single-end, 50-base reads for miRNA sequencing.