GSM7059907: Dyak.ovary.3 small-RNA-Seq; Drosophila yakuba; ncRNA-Seq
Small RNAs were isolated using the TraPR Small RNA Isolation Kit (Lexogen) following the manufacturer's instructions. The soma-enrichment sRNA-seq libraries were generated for 13 species (2 replicates each) similar to published protocols (Akkouche et al., 2013; Chirn et al., 2015), with modifications.  In brief, 75-100 ovary pairs were dissected in ice-cold PBS.  Ovaries were dissociated for 18 minutes in 0.25% Trypsin (Sigma-Aldrich) at 25˚C, shaking at 800 rpm.  Dissociated tissue was pushed through a 40 µm nylon mesh (Greiner Bio-one) washed with equal volume Schneider 2 medium (Thermo Fisher Scientific) and then pelleted.  Cell count was quantified using the Luna-FL (Logos Biosystems).  Cells were pelleted and directly used as input for sRNA isolation using the TraPR Small RNA Isolation Kit (Lexogen). sRNA libraries were generated using the Small RNA-Seq Library Prep Kit (Lexogen) with minor modifications.  Both primers A3 and A5 as well as the primer RTP were used at 0.5x.