GSM7081673: Sperm_ChIP_H3K4me3_DTT; Mus musculus; ChIP-Seq
Sperm were first fixed with 1% formaldehyde in PBS for 10 min at room temperature, followed by quenching with Glycine. Next, sperm were pre-treated with 50 mM DTT and 1 mg/ml heparin at 37°C for 2 hr. Sperm were then quenched with NEM and washed with PBS twice.  After lysing sperm in complete buffer containing detergent, equal volume of 2x RIPA buffer was supplied to make a 1x RIPA condition. Chromatin shearing was performed using Covaris S220 platform with parameters: peak power 140 W, duty factor 5% and 200 Cycles/burst for a total of 30 min. Sonicated sperm chromatin were directly subjected to immunoprecipitation with corresponding antibodies Library preparation was carried out using NEBNext Ultra II DNA Library Prep kit (NEB E7645)