GSM7755829: Sample 12_ATA12, WT; Mus musculus; ATAC-seq
ATAC-seq were carried out following the Omni-ATAC-seq protocol (Corces et al., 2017). Briefly, 50 k cortical microglia (CD11b+Fcrls+ Ly-6C–) from Itgb8fl/fl;Emx1Cre mouse brains cells were sorted into Eppendorf tubes by a BD FACSAriaTM II (BD Bioscience). After being washed with cold dPBS, microglia were permeabilized with ATAC-Resuspension Buffer containing 0.1% NP40, 0.1% Tween-20, and 0.01% Digitonin and washed with ATAC-RSB buffer with 0.1% Tween-20. The isolated microglial nuclei were then incubated in transposition mixture at 37℃ for 60 min. The DNA were purified using a Zymo DNA clean and concentrator-5 kit (Zymo research, Cat.#D4014) and amplified using NEBNext® High-Fidelity 2x PCR Master Mix (NEB, Cat. #M0541s) for 9 cycles. The ATAC libraries were size selected using SPRIselect beads (Beckman Coulter, Cat.#B23317) and quantified using Agilent 2100 Bioanalyzer and Qubit (Invitrogen, Cat.#Q33238). The pooled libraries were then sent to Azenta Life Sciences for paired-end sequencing (2x150 bp) on Illumina HiSeq platform.