title: GSM8854931: BAT, OE Runx2, HDAC1 CUT&Tag, replicate 1; Mus musculus; OTHER
library_construction_protocol: For ChIP-seq, brown preadipocytes or brown adipocytes were fixed with 1% formaldehyde for 10min and the reaction was quenched by 0.125M glycine. Cells were then washed with PBS for two times. Cells were lysed in ChIP buffer (1% SDS, 50 mM Tris-HCl, pH 8.0, 10 mM EDTA, and protease inhibitor cocktail) for 30 min at 4°C. The DNA was fragmented to 200-500bp by sonication and centrifuged at 12,000 g for 5min. The supernatant was diluted with ChIP IP buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl and protease inhibitor cocktail). For the CUT&Tag assay was performed using the NovoNGS® CUT &Tag 2.0 A High-Sensitivity Kit (for Illuina®) (Cat# N259-YH01-01A, Novoprotein Scientific, Inc.,) following the manufacturer's protocol. The ChIP DNA library was constructed with VAHTS Turbo DNA Library Prep Kit for Illumina (Vazyme Biotech) according to the manufacturer's instructions. For the CUT&Tag assay was performed using the NovoNGS® CUT &Tag 2.0 A High-Sensitivity Kit (for Illuina®) (Cat# N259-YH01-01A, Novoprotein Scientific, Inc.,) following the manufacturer's protocol.