GSM4984038: pupa12h_brain_DGRP_mix_DGRP-551_Hyb-409-502-639_sample2_seq1; Drosophila melanogaster; ATAC-seq
D. melanogaster adult brains were dissected and transferred to a tube containing 100 µl ice cold DPBS solution. After centrifugation at 800 g for 5 min, the supernatant was replaced by 500 µl nuclei lysis buffer composed of 10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% Nonidet P40, 0.01% Digitonin, and 1% BSA, in Nuclease-free water. The following procedure was followed to extract the nuclei from the brain tissue: incubation in nuclei lysis buffer on ice for 5 min, transfer to a dounce tissue grinder tube (Merck), 25 strokes with pestle A, incubation on ice for 10 min, 25 strokes with pestle B. The lysis was stopped by adding 1 ml of wash buffer composed of 10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20 and 1% BSA, in Nuclease-free water. Nuclei were pelleted by centrifugation at 800 g for 5 min at 4°C and resuspended in 1x Nuclei Buffer (10x Genomics). Nuclei suspensions were passed through a 40 µM Flowmi filter (VWR Bel-Art SP Scienceware). Nuclei concentration was assessed by the LUNA-FL Dual Fluorescence Cell Counter. 10x Genomics CHROMIUM Single Cell ATAC Library & Gel Bead Kit v1