title: GSM6923713: Hippocampus, ATAC, cKO, rep2; Mus musculus; ATAC-seq
library_construction_protocol: P0 hippocampi were dissected. and gently homogenized in cold nuclear isolation buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Igepal CA-630). The nuclei are collected by centrifugal precipitation. ATAC-seq libraries were constructed by TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme TD501). 50,000 nuclei were put into the tagmentation reaction for each sample (performed with 30 min incubation time at 37°C). Immediately following the tagmentation, we purified DNA fragments using VAHTS DNA Clean Beads (2X). We took purified DNA fragments and added Illumina i5+i7 adapters with unique index to individual samples by PCR reaction (PCR program: 72°C for 3 min, 98°C for 30 s, 98°C for 15 s, 60°C for 30 s, 72°C for 30 s, repeat 3-5 for 13 cycles, 72°C for 5 min, and hold at 4°C). Generated libraries were purified using VAHTS DNA Clean Beads (1.2X). For multiplexing, libraries with different barcode were mixed together with equal molar quantities by considering appropriate sequencing depth (about 50 million reads per library). Libraries were sequenced by Illumina Nova-seq 6000 platform with pair-end reads of 150 bp.