GSM5460652: control knockdown (GFP) in mature gamergate - rep 3; Harpegnathos saltator; RNA-Seq
Harpegnathos ovaries were dissected from single individuals, snap-frozen and then homogenized in TRIzol (Thermo Fisher Scientific, MA). RNA was purified and its quality visualized on agarose-formaldehyde gels. For library preparation, polyA+ RNA was isolated from total RNA using Dynabeads Oligo(dT)25 (Thermo Fisher) and constructed into strand-specific libraries using the dUTP method (Parkhomchuk et al., 2009). UTP-marked cDNA was end-repaired using end-repair mix (Enzymatics, MA), tailed with deoxyadenine using Klenow exo- (Enzymatics), and ligated to custom dual- indexed adapters with T4 DNA ligase (Enzymatics). Libraries were size-selected with SPRIselect beads (Beckman Coulter, CA) and quantified by qPCR after amplification.