title: H3K4me3 ChIP-seq of mus musculus: adult male PFC
design: Chromatin immunoprecipitation sequencing (ChIP-seq) was performed on prefrontal cortex tissue from control, CSDS10, and CSDS30 groups of C57BL/6 mice. Animals were euthanized 24 hours after the last social defeat stress exposure. Prefrontal cortex samples were dissected and stored at -70C prior to processing. Chromatin was fragmented using micrococcal nuclease (MNase), which selectively digests linker DNA while preserving nucleosome-bound DNA. Approximately 20 mg of tissue per sample was incubated overnight at 4C with anti-H3K4me3 antibodies (ab8580, Abcam). Immunoprecipitated complexes were eluted and DNA was purified using phenol-chloroform extraction. Libraries were prepared using NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer protocol. DNA fragment size selection was performed with AMPure XP beads (Beckman Coulter), followed by PCR enrichment.