title: GSM5508126: HeLa_Input; Homo sapiens; ChIP-Seq
library_construction_protocol: To extract DNA, the pull-down chromatin fractions were incubated in de-crosslinking buffer (50 mM Tris-HCl pH 8.0, 200 μg/mL Protease K, 2% SDS) overnight at 65°C. An equal volume of phenol: chloroform: isoamyl alcohol (25:24:1,v/v) (311-90151, Wako) was added and vortexed, and centrifuge at 16,000 g for 5 min at room temperature. The upper aqueous phase was transferred to a new tube. This step was repeated. One-tenth volume of 3 M sodium acetate, glycogen, and the double volume of ethanol were added and incubated for 2 hr at -80°C. After centrifugation at 20,000 g for 30 min at 4ºC, the pellet was retained, washed with 70% ethanol, and dried up. The dried pellet was dissolved in 10 μg/mL RNase A in DNA dilution buffer attached to DNA SMART ChIP-Seq Kit (634865, Takara). After incubation for 1 hr at 37°C, DNA was purified by the phenol-chloroform extraction and ethanol precipitation to remove RNase A. DNA libraries were synthesized by the DNA SMART ChIP Seq Kit (634865, Takara).