WGS Metagenomics of Drosophila melanogaster Oregon-R-modENCODE, Adult,  Female, Guts
bacterial DNA was isolated from 20 adult 21-day old female fly guts.  Whole flies were briefly anesthetized with CO2, washed once with 3% sodium hypochlorite and once with 70% ethanol, then dissected in sterile 1xPBS.  The dissected guts were homogenized in 100uL 1xPBS buffer using a motorized pellet pestle with a disposable, autoclaved pestle.  Following centrifugation at 1780 x g for 3 minutes, the supernatant was discarded and the pellet was resuspended in 100uL of QuickExtract DNA extraction solution (QE09050,  Lucigen) and 1L ReadyLyse Lysozyme Solution (30,000 U/L, R1804, Lucigen).  DNA was fragmented using NEBNext dsDNA Fragmentase (M0348S, New England BioLabs) and EDTA was removed with 1.5 volumes of AMPure XP (A63882, Beckman-Coulter Life Sciences) beads.  Libraries were made using NEBNext Ultra DNA Library Prep Kit for Illumina (E7370S, New England BioLabs) and sequenced using a MiSeq v2 Reagent Kit (MS-102-2002, Illumina).