RNA-Seq Drosophila melanogaster virgin  female abdomen
Male and female flies were either kept as virgins, or mated once and flash frozen 3-hours post-mating. Pools of 50 flies per sample were separated into head-thorax and abdomen body-parts and the mRNA and sRNA fractions were extracted using the mirVana miRNA isolation kit (Ambion) according to manufacturers instructions. Samples were DNase treated to remove residual genomic DNA (Ambion Turbo DNA-free kit). RNA concentration and integrity was measured on the NanoDrop 8000 Spectrophotometer (Thermo Fisher Scientific). 32 libraries (two replicates, two sexes, two body-parts, two treatments, two RNA types) were constructed by the Earlham Institute, Norwich Research Park using the Illumina TruSeq kit. For the sRNA libraries, a modified blocking oligo was also used to preclude adapter ligation to the highly abundant 30 nt 2S rRNA. Non-directional, single end RNA-seq was conducted using the Illumina HiSeq2500 platform with 50nt read length.