title: GSM6249134: KP4-DMSO-Input-rep-1; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP lysates were prepared from 2X10^7 cells. Cells were fixed in 1% formaldehyde for 10min and lysed for 10min. The lysed cells were sonicated using Covaris Focused Sonicator at 4°C. Chromatin was subjected to ChIP using 3µg of antibody and incubated overnight at 4°C. Protein A Dynabeads was added to the chromatin and incubated for 2 hours, followed by sequential washing with ice-cold RIPA buffer containing 150 mM salt (50mM Tris-HCl pH 8, 0.15M NaCl, 1mM EDTA pH 8, 0.1% SDS, 1%  Triton X-100, 0.1% sodium deoxycholate), 500 mM salt (50mM Tris-HCl pH 8, 0.5M NaCl, 1mM EDTA pH 8, 0.1% SDS, 1% Triton X-100, 0.1% sodium deoxycholate), twice with lithium chloride buffer (50mM Tris-HCl pH 8, 1mM EDTA pH 8, 1% Nonidet P-40, 0.7% sodium deoxycholate, 0.5M LiCl2) and 1xTE buffer (10mM Tris-HCl pH 8, 1mM EDTA pH 8). DNA was eluted for 4h at 55°C and reverse crosslinked overnight with Proteinase K at 65°C followed by phenol/chloroform extraction and ethanol precipitation. Eluted DNA was used for library preparation. Libraries were prepared using Illumina DNA Flex library preparation method following the manufacturer's protocol.