RNA-Seq of Drosophila melanogaster: adult female ovary
Each replicate of 10 pairs of ovaries remained frozen until total RNA was isolated from them using the Zymo Direct-zol kit. Integrity of the isolated RNA was evaluated using an Agilent Fragment Analyzer, and RQN values ranged from 8.9-10. Poly-A-selected mRNA-seq libraries were prepared using the NuGen (Tecan) Universal Plus kit, which introduces chemical fragmentation of the mRNA prior to first strand synthesis. RNA isolations and RNA-seq libraries were prepared and sequenced at the University of Oregon Genomics and Cell Characterization Core Facility. Libraries were sequenced on a HiSeq 4000 using single end, 150-bp reads.