title: GSM8144900: GSC1228_D_K4m1; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells (1 × 106) were treated with 1% formaldehyde for 8 min to crosslink histones to DNA. After washing with cold PBS, cell pellets were lysed with lysis buffer (10 mM Tris-HCl, 0.25% Triton X-100, 10 mM EDTA, and 0.1 M NaCl). Cell nuclei were collected after centrifugation and re-suspended in SDS lysis buffer (1% SDS, 150 mM NaCl, 50 mM HEPES-KOH, 2 mM EDTA, 1% Triton X-100, and 0.1% sodium deoxycholate). Chromatin was sonicated using Covaris S220. Lysates were incubated overnight with 50 μl of Dynabeads Protein G which were pre-coated with antibodies. The beads were washed and protein-DNA complexes were eluted with elution buffer (1% SDS, 10 mM EDTA, and 50mM Tris-HCl) and then treated with RNase for 2 h at 37°C followed by proteinase K treatment overnight. DNA was extracted by the phenol/chloroform method, ethanol-precipitated and resuspended in 0.1 × TE buffer. 10% of the lysate was treated with RNase and proteinase K and de-crosslinked as input samples. NEBNext Ultra II DNA library Prep kit