GSM9554032: R65H02, Tempearture 29, Gal4+, Control, ZT0, Thermogenetic Sleep Deprivation,rep-3; Drosophila melanogaster; RNA-Seq
At the appointed time (usually immediately after sleep deprivation at ZT0 or immediately after sleep promotion at ZT12), flies were removed from the incubator and brains were dissected in dissecting saline with channel blockers. Likewise, circadian profiling occurred in the same way at the appropriate time point. 15 brains per sample were pooled, transferred to lysis buffer from the PicoPure RNA Isolation kit, vortexed for 30 seconds, then incubated at 42C for 30 min before freezing. Once all samples were collected, the RNA was isolated using the PicoPure RNA Isolation kit and used for library prep. Libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit (Cat #: E7770) following manufacturer instructions.