GSM4891862: Dueling gamergate brain - d10 - rep4; Harpegnathos saltator; RNA-Seq
Brains were harvested by dissection in neurobasal medium, the central brain region was isolated by removing the optic lobes, gnathal ganglion, and ~95% of the antennal lobes dissected in neurobasal medium (Gibco), and the tissue was then processed for RNA- or ChIP-seq (see below). 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.