title: GSM4684704: MCF7_Myc_ChIPseq; Homo sapiens; ChIP-Seq
library_construction_protocol: For ChIP-seq DNA extracton, 1.2×107 cells were treated with 5 mM dimethyl 3,3'- dithiobispropionimidate (Thermo Scientific) for 30 min at 4°C and incubated with quenching buffer (100 mM Tris-HCl, 150 mM NaCl) for 5 min. Treated cells were cross-linked with 1% (vol/vol) formaldehyde for 10 min at room temperature, quenched with 125 mM glycine for 5 min, lysed in lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl, pH 8.0), and sonicated by the Bioruptor (Diagenode) to generate DNA fragments ranging from 200 to 600 bp. The sonicated lysates were diluted with ChIP dilution buffer (0.1% TritonX-100, 2 mM EDTA, 20 mM Tris-HCl pH 7.5, 150 mM NaCl, and 1× protease inhibitor), immunoprecipitated with rotation 6-7 hours at 4°C by 3-4 µg of anti-MYC antibodies, pulled down using Protein A/G DYNAL magnetic beads (40 µl of 1:1 mixture), and eluted by the QIAquick PCR Purification Kit (Qiagen). ChIP-seq library preparation and sequencing were performed by a NextSeq500 platform with single-end reads of 75 bases
attributes.tag: GEO Accession
attributes.value: GSM4684704