GSM8111955: Repo_ZT14_1; Drosophila melanogaster; ATAC-seq
ATAC-Seq was performed on FACS-sorted clock neurons and glia using Clk856-Gal4 and Repo-Gal4, crossed to UAS-unc84-GFP, respectively. Brains were dissected in Schneider's media and incubated with 0.75mg/ml Collagenase and 0.4mg/ml Dispase at room temperature for 30 minutes. Brains were then triturated 50-80 times to obtain a single-cell suspension for FACS sorting. Sorted cells and nuclei were centrifuged at 1000 rpm for 10mins at 4°C and the pellet was resuspended in 50uL of Transposition mix (25uL 2xTD buffer, 16.5uL PBS, 0.05 μl 10% v/v Tween, 0.05 μL 1% v/v Digitonin, and 2.5uL TDE1 Enzyme (Illumina, San Diego, CA, USA Catalog #20034198) and incubated at 37 °C for 30 mins. Tagmented DNA was purified using Zymo DNA clean and concentrator kit. Purified DNA fragments were amplified using single-indexed Nextera primers (Integrated DNA Technologies) for 12 PCR cycles. Amplified libraries were purified using AMPure XP beads (Beckman Coulter, Brea, CA, USA Catalog #A63880), and size distribution was accessed using TapeStation High-Sensitivity D1000 Screentape. Final libraries were sequenced on a Nextseq 550 (Illumina).