D. simulans w[XD1] RNA-seq replicate 2
We extracted RNA from testes (dissected free of accessory glands) of 7-day-old w[XD1] male flies using Trizol (Invitrogen). Illumina Truseq Total RNA library Prep Kit LT was used to make stranded RNA-seq libraries from 650 ng of total RNA. Manufacturers protocol was followed except for using 8 cycles of PCR to amplify the final library instead of the recommended 15 cycles, to minimize artifacts caused by PCR amplification. All samples were pooled together using the barcoded adapters provided by the manufacturer and the sequencing was performed by HiSeq2500 instrument using PE75 at the New York Genome Center.