GSM7755845
Anti H3K9ac ChIP-seq were performed using using the iDeal ChIP-seq kit for Histones (Diagenode, Cat.#C01010059) and NEBNext® UltraTM II DNA Library Prep Kit for Illumina® (NEB, Cat.#E7103S). Briefly, microglia were sorted from Itgb8fl/fl;Emx1Cre mouse brain cortex and were pooled from different individuals with same genotype to achieve > 200 k cells per replicates. The microglia were fixed with 1% formaldehyde at 20℃ for 8 min then quenched with 0.125 M Glycine. The nuclei were isolated and sonicated using a Diagenode disruptor for 20 cycles (30 seconds “ON”, 30 seconds “OFF”, high power). The chromatin fragments were enriched using 2 ul of anti-H3K9ac antibody (Millipore, 07-352) per reaction. The ChIPed DNA was purified and processed for library preparation. The ChIP DNA 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.