title: GSM8824285: mm_penis_P13_ARChIP; Mus musculus; ChIP-Seq
library_construction_protocol: We placed dissected tissue in ice cold 1x DPBS for ChIP-seq. We incubated penis, forelimb, and hindlimb tissues in Trypsin-EDTA for 5 minutes at 37°C to loosen up the tissue. We inactivated Trypsin-EDTA using DMEM + 10% FBS, then cross-linked DNA to bound proteins by adding 1% formaldehyde, incubated on a horizontal nutator for 20 minutes at room temperature. We quenched the crosslinking reaction using 2.5 M Glycine, spun down tissues at 4,000 rpm for 5 minutes at 4°C. We washed tissues twice in ice-cold PBS, then removed the tissue pellet. We sheared chromatin to a size of 100-300 bp in a bath-based sonicator (Diagenode Bioruptor 300). We produced ChIP and input chromatin control libraries using the NEBNext Ultra II Library Prep Kit (NEB, catalog #E7645S).