GSM9017659: PC3_SMARCD2; Homo sapiens; ChIP-Seq
Crosslinked cells were lysed and sonicated as described previously (K. A. Holmes, G. D. Brown, J. S. Carroll, Methods Mol. Biol., 2016). Briefly, crosslinked cells were lysed and nuclei were enriched sequentially in Lysis buffer 1 (LB1: 50 mM HEPES—KOH, pH 7.5, 140 mM NaCl, 1 mM  EDTA, 10% glycerol, 0.5% Igepal CA-630, 0.25% Triton X-100), Lysis buffer 2 (LB2: 10 mM Tris—HCl, pH 8.0, 200 mM NaCl, 1 mM EDTA, 0.5 mM EGTA), and Lysis  buffer 3 (LB3: 10 mM Tris—HCl,  pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 0.1% Na-deoxycholate, 0.5% N-lauroylsarcosine). Chromatin was fragmented using 4 minutes of active sonication (30 sec ON 60 sec OFF). 5 ng input and IP DNA was used for sequencing library construction using NEBNext Ultra II DNA Library Prep Kit for Illumina (Cat # E7645, NEB) according to the manufacturer's instructions.Libraries were amplified using KAPA HiFi HotStart Ready Mix (Roche, Cat # KK2602) for 10 cycles. Amplified libraries were purified using Agencourt AMPure XP Beads (Beckman Coulter, Cat # A63881).