GSM8604691: male, chow diet, sample 486; Mus musculus; ATAC-seq
Nuclei were isolated from skeletal muscle of normal chow-fed, Control f/f and mERaOE mice at months of age (n=3/genotype). Briefly, 50 mg of frozen gastrocnemius muscle was minced and dounced in NIB buffer (20mM Tris-HCl, 50mM EDTA, 5mM Spermidine, 0.15mM Spermine, 0.1% mercaptoethanel, 40% glycerol pH=7.5) for 120 strokes. Homogenize samples were incubated on ice for 5 min and centrifuged at 11,00g at 4°C. The supernatants were filtered through 70 μm nylon mesh and washed with RSB buffer (10mM Tris-HCl, 10mM NaCl, 3mM MgCl2 pH7.4). ATAC-seq libraries were prepared from 50,000 nuclei using the Nextera Tn5 Transposase and DNA library preparation kit (Illumina) as described with slight modifications