title: GSM6659480: +4OHT Universal 2% Input Control; Mus musculus; ChIP-Seq
library_construction_protocol: ChIP was performed using the SimpleChIP® Plus Enzymatic Chromatic IP with Magnetic Beads Kit (Cell Signaling) protocol according to manufacturers instructions. In short, after 48h in either DMSO or 1μM 4OHT, media was removed and monolayers were cross-linked for 20 minutes in 1% PFA. Following fixation quenching with glycine and pelleting, nuclei were isolated, treated with micrococcal nuclease, and sonicated four times for 15 seconds each with an ultrasonic sonicator with a 1/8-inch probe on ice to shear chromatin into 150bp-900bp fragments. 5ug of cross-linked, digested chromatin was immunoprecipitated using 1ug of either rabbit anti-p63α (Cell Signaling, clone D2K8X), rabbit anti-H3K27ac (Cell Signaling, clone D5E4), or rabbit anti-H3K27me3 (Cell Signaling, clone C36B11). ChIP-seq libraries were prepared using using the NEBNext® UltraTM II DNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions. After library cleanup, library concentrations were quantified using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific) and size validated on a TapeStation (Agilent Technologies).