GSM1973165: 4096; Ovis aries; RNA-Seq
EVs were isolated from culture media by adding 200 µl ExoQuick-TC (System Biosciences) precipitation solution per ml of filtered (0.2 µm) media per the manufacturer's instructions. Total RNA from day 14 conceptus EVs isolated from conceptus-conditioned culture media (n=3) was purified using the miRNeasy kit (Qiagen, Valencia, CA). RNA concentration was determined by quantitative high sensitivity RNA analysis on the Fragment Analyzer instrument (Catalog # DNF-472, Advanced Analytical Technologies, Inc., Ankeny, IA). Libraries were constructed following the manufacturer’s protocol with reagents supplied in Clontech SMART-Seq® v4 Ultra™ Low Input RNA Kit for Sequencing kit (Catalog # 634888) and Low Input Library Prep Kit (Catalog # 634947, Clontech Laboratories, Inc., Mountain View, CA) according to the manufacturer’s instructions using 5 ng of total RNA. Libraries were diluted and sequenced according to Illumina’s standard sequencing protocol on a HiSeq 2500 (paired-end 50 bp).