GSM2500284: EGFP::ASH1DBAH Rep 2; Drosophila melanogaster; RNA-Seq
Total RNA was extracted from dissected 3rd instar larval wing discs using Trizol Reagent (Ambion). RNA was additionally treated with DNase (Turbo DNase, Ambion). cDNA was synthesized using the SuperScript II kit (Invitrogen) according to the manufacturer's instructions. Priming was performed with equal amounts of oligo(dT) and random decamer primers (Ambion). RNA was removed by treatment with RNase H (NEB) before PCR analysis or mRNA purification and sequencing. mRNA was purified from total RNA using Oligo (dT)25 Dynabeads (Invitrogen), fragmented by heat treatment and converted to cDNA using Superscript III (Invitrogen). Sequencing libraries were prepared with the KAPA Library Preparation Kit Illumina series (KK8201). Libraries were sequenced on an Illumina HiSeq 2000.