GSM2775048: 4165 miRNA-Seq; Ovis aries; miRNA-Seq
Endometrium was physically dissected from the uterus at collection and snap frozen in liquid nitrogen. Total RNA was isolated from tissue homogenzied in TRIzol reagent. The uterine lumen was flushed with 10 ml sterile PBS. Recovered uterine flush was clarified by centrifugation, filtered through a 0.22 µm PVDF syringe filter and concentrated by ultrafiltration. Extracellular vesicles (EVs) were isolated from concentrated flush by size exclusion chromatography (SEC) using qEVoriginal columns (Izon Science Ltd, Oxford, United Kingdom). Total RNA was isolated from EVs with the miRNeasy kit including on-column DNase digestion. Illumina TruSeq small RNA