GSM7870940: E9.5M_162_Input; Mus musculus; ChIP-Seq
Chromatin from hemisected placental tissues were fixed with 1% formaldehyde rotated for 12 minutes at room temperature and was subsequently quenched using a final concentration of 125mM glycine. Samples were thoroughly homogenized and washed with ice cold PBS. Fixed chromatin was sonicated using a Covaris E220 for 30-60 minutes at 4°C with the following conditions: peak incident power, 140; duty factor, 10%; Cycles/burst, 200; Water level, 0. Equal amounts of chromatin per sample were rotated with select antibodies (2.5 μg antibody/sample of either H3K4me3Q5ser (MilliporeSigma ABE2580) or H3K4me3 (Active Motif 39159)) bound to M-280 Dynabeads at 4°C overnight. The next morning, samples were washed, eluted, and reverse-crosslinked at 65°C. Samples underwent RNA and protein digestion, and DNA was purified using QIAQuick MinElute Spin columns (Qiagen 28140). 1% inputs were removed prior to antibody incubation and purified in parallel with corresponding immunoprecipitates. TruSeq ChIP Library Preparation Kit (Illumina IP-202-1012)