title: D_simulans_males_R1
design: To collect RNA sequencing data, D. simulans (wXD1), D. mauritiana (w12) and D. sechellia  (Rob3c / Tucson 14021-0248.25) were reared at room temperature in vials on standard cornmeal media or standard cornmeal-molasses medium. Males and virgin females were collected post-eclosion and allowed to age for 3-5 days in single-sex vials.  20-30 flies were then flash frozen in liquid nitrogen and stored at -80C for RNA extraction. Testes from at least 100 males were dissected in PBS buffer and stored at -80C. For D. simulans and D. mauritiana, Total RNA was extracted following standard protocols using Trizol (Invitrogen), chloroform, and phase-lock gel tubes (Fisher Scientific).  Sequencing libraries were generated using the Illumina TruSeq Stranded mRNA kit with 125 bp inserts and paired-end reads were sequenced on an Illumina HiSeq 2500 at the University of Minnesota Genomics Center.