GSM5345080: SETMAR-BS_input; Homo sapiens; ChIP-Seq
For ChIP, 2x107 cells were suspended in 1.5ml of ChIP Lysis Buffer (50mM HEPES-KOH pH 7.5, 140mM NaCl, 1mM EDTA pH8, 1% Triton X-100, 0.1% Sodium Deoxycholate, 1% SDS, supplemented with protease inhibitors) and disrupted by Fast prep (speed: 6 m/s; time: 40 sec; 2 cycles; pause time: 120 sec). Chromatin lysates were sonicated in 1.5 ml LowBind  tubes (600 μl sample/tube using Diagenode Bioruptor Plus (2x5 cycles, 30 sec ON/OFF LOW). After sonication, cell debris were pelleted by centrifugation at 16.000 g (at 4°C for 20 minutes). Fragment length distribution of sonicated samples (50 μl) was checked by 1% agarose gel electrophoresis after reverse crosslinking (at 65 °C for 6 hours) and phenol-chloroform extraction. NGS libraries were prepared by the Nugen Ovation Ultralow System V2 library preparation kit (NuGEN Technologies) following the manufacturer's instructions.
GEO Accession
GSM5345080