GSM8726787: supine_study_Age_62_Surv_37_6_Rep1; Drosophila melanogaster; RNA-Seq
Individual flies were homogenized in Lysis Binding Mix (from MagMAX mirVana Total RNA Isolation Kit; Thermo Fisher Scientific, Washington, DC; #A27828) using Navy Eppendorf RNA Lysis Kits (Next Advance, Troy, NY; #NAVYE5-RNA) and a Bullet Blender Storm 24 (Next Advance, Troy, NY; #4116-BBY24M). RNA was then isolated from these homogenates using the MagMAX mirVana Total RNA Isolation Kit as described in the manufacturer's instructions, in conjunction with the KingFisher Apex Purification System with 96 Deep-Well head (Thermo Fisher Scientific, Washington, DC; #5400930). RNA quality and quantity was first assessed by NanoDrop (Thermo Fisher Scientific, Washington, DC) and High Sensitivity RNA ScreenTape (Agilent Technologies, Santa Clara, CA). cDNA libraries were generated from 500 ng of total RNAusing NEBNext Ultra II Directional Poly-A RNA Library Prep Kit for Illumina (New EnglandBiolabs, Ipswich, MA; #E7760). cDNA library quality and quantity were assessed byTapeStation 2200 and Qubit (Thermo Fisher Scientific, Washington, DC). cDNA librarieswere sequenced on a NextSeq 2000 machine using NextSeq 2000 P3 reagents (Illumina,San Diego, CA).