GSM4891916: 20E baseline - rep 1; Harpegnathos saltator; RNA-Seq
Cells were lysed in TRIzol (Thermo Fisher Scientific, MA). RNA was purified and its quality visualized on agarose-formaldehyde gels. Typical yields amounted to 0.6 µg total RNA per central brain. For library preparation, polyA+ RNA was isolated from 600 ng 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.