title: GSM8877188: white_H3K9me3_D15; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were fixed for 10 min with 1% formaldehyde, lysed in 50 mM Tris-HCl, pH 8, 10 mM EDTA, 1% SDS, protease inhibitors and Na-butyrate, and sonicated in a Picorupter (Diagnode) into ~200 bp fragments. After sedimentation, the supernatant was diluted 10 times in RIPA buffer and incubated with H3K4me3 (Diagenode c15410003), H3K4me1 (Diagenode c15410037) or H3K27ac (Diagenode c15410174) antibodies (each at 2.5 µg/106 cells) coupled to magnetic Dynabeads Protein A/G (Invitrogen) for 2 h at 4°C. ChIP samples were washed, cross-links were reversed and DNA was eluted for 2 h at 68°C in 50 mM NaCl, 20 mM Tris-HCl pH 7.5, 5 mM EDTA, 1% SDS and 50 ng/µl Proteinase K. DNA was purified using phenol-chloroform isoamylalcohol and dissolved in H2O. For Day 15 differentiated white and beige adipocytes, cells were trypsinized, resuspended in HBSS, 0.5% BSA, and centrifuged 200g for 5 min to isolate floating mature adipocytes. Purified nuclei were then fixed and processed as cell samples ChIP-seq libraries were prepared using a Microplex kit (Diagenode) and sequenced with 150-bp paired-end reads on an Illumina NovaSeq.
identifiers.namespace: GEO
identifiers.id: GSM8877188_r1