title: GSM8879160: FACS_sorted_Ms_Purkinje_neurons_NT_40_ATAC_seq; Mus musculus; ATAC-seq
library_construction_protocol: For ATAC-seq, the nuclei pellet was resuspended in 190μl transposition reaction mix (10mM Tris-HCl pH 7.5, 5mM MgCl2, 10% Dimethylformamide) by pipetting up/down 10 times. 10μl of pre-assembled Tn5 transposome with ATAC adapters was added. Tubes were gently agitated to mix all components and incubated at 37°C for 30 minutes at 1200 rpm in an Eppendorf Thermomixer. ATAC reactions were terminated by adding 1mL Zymo DNA binding buffer and vortexed. Reactions were purified according to the manufacturer's instructions using the DNA Clean and Concentrator-5 kit (ZYMO RESEARCH, cat. No. D4004) and eluted in 15μl nuclease-free water. For ATAC-seq, eluted ATAC DNA was gap repaired, purified by the DNA Clean and Concentrator-5 kit, and eluted in 25μl nuclease-free water. Eluted ATAC DNA was amplified and barcoded in 50μl PCR reactions (25μl 2x Kapa HiFi HotStart Ready Mix, 22μl eluted ATAC DNA, 1.5μl 10μM i5 index primer, 1.5μl 10μM i7 index primer) with the following PCR thermocycler program: 98°C 45 s; 10 cycles of 98°C 15 s, 62°C 30 s, 72°C 30 s; final extension 72°C 2 min; hold at 4°C. PCR reactions were cleaned by AMPure XP beads.
identifiers.namespace: GEO
identifiers.id: GSM8879160_r1