GSM7946841: 3rG4 cl6 k27me3 rep1 input; Mus musculus; ChIP-Seq
10 million cells per ChIP were resuspended with 0.5 ml ice-cold ChIP lysis buffer (50 mM Tris-HCl pH8, 10 mM EDTA pH8, 1% SDS).  Cell extracts were subjected to sonication using the Qsonica apparatus (40% power, 30s on/ 30s off for total sonication time 30 min at 4°C). Sheared chromatin was checked on the 1% agarose gel electrophoresis, repeat sonication a second time if the fragment is larger than 1 kb marker. 0.1 ml lysate was mixed with 1.2 ml ChIP dilution buffer (20 mM Tris-HCl pH8, 150 mM NaCl, 1% Triton X-100, 2 mM EDTA pH8), and incubated with 2 μg antibody for overnight at 4°C. Immunoprotein complex was captured with 20 μL Dynabeads Protein G for 5 hr incubation. Afterward, beads were washed three times with ChIP wash buffer 1 (20 mM Tris-HCl pH8, 150 mM NaCl, 2mM EDTA pH8, 1% Triton X-100, 0.1% SDS) and once with ChIP wash buffer 2 (20 mM Tris-HCl pH8, 500 mM NaCl, 2mM EDTA pH8, 1% Triton X-100, 0.1% SDS). 100 μL of ChIP elution buffer (1% SDS, 100 mM NaHCO3, 40 ug/ml RNaseA) was added to the washed bead and incubated for 1hr at 37°C to recover bound nucleoproteins, dynabeads were removed by using magnetic stand. Reverse-crosslinking was done by incubating the supernatant at 65° C overnight. NEBNext Ultra II DNA Kit (NEB) was used for library preparation by following the product manual.