Instrument: Illumina HiSeq 2500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Brains were dissected out, flash frozen, and stored at -80C until sectioning on a cryostat at -20C for tissue-punching of the medial prefrontal cortex. Chromatin was then crosslinked and sheared to fragments of 200-500 bp using a Bioruptor sonicator (Diagenode, Denville, NJ, USA). Next, pre-cleared chromatin was immunoprecipitated overnight at 4℃ with an antibody directed against Egr1 (sc-110-X, 4 μg, Santa Cruz Biotechnology, Dallas, TX, USA). After washes, elution from beads, and reversal of the cross-link, immunoprecipitated DNA was purified. ChIP-Seq libraries were prepared using the NEBNext Ultra DNA Library Prep Kit for Illumina and barcoded primers (New England Biolabs) following the manufacturer’s protocol (#E7370). For each biological sample, two libraries were prepared (IP and Input), resulting in a total of 22 barcoded libraries.