title: GSM4981347: 293T_ZNF676_rep3; Homo sapiens; ChIP-Seq
library_construction_protocol: 30 million cells per IP were collected and the chromatin was prepared and sonicated as described (Imbeault et al., 2017).  Total input samples (one per cell line) were collected. The IP was then performed with the anti‐HA.11 antibody (clone 16B12, Covance, MMS-101P) incubated overnight at 4°C on a rotating wheel, and captured using protein G Dynabeads (Thermo Fisher, 10004D). Samples then underwent 5-minute washes on a rotating wheel: twice in low salt buffer (10 mM Tris-HCl pH 8, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.15% SDS, 1x PMSF), once in high salt buffer (10 mM Tris-HCl pH 8, 500 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.15% SDS, 1x PMSF), once in LiCl wash buffer (10 mM Tris-HCl pH 8, 1 mM EDTA, 0.5 mM EGTA, 250 mM LiCl, 1% NP40, 1% NaDOC, 1x PMSF). TE buffer was then added to the beads and the supernatant was removed.  Beads were resuspended in elution buffer (TE buffer, 1% SDS, 150 mM NaCl), treated with Proteinase K (Thermo Fisher, AM2548) and decrosslinked overnight at 65°C and 1100 rpm. The DNA was then purified using a MinElute PCR Purification kit (Qiagen, 28004). NextSeq v2.5 (with Illumina protocol #15048776)