GSM5662052: Utx - KO [H3K27me3 CUT&Tag]; Mus musculus; OTHER
Muscle satellite stem cells were isolated from indicated muscles by mincing and straining, which were then sorted by fluorescence activated cell sorting. FACS sorted cells were collected in microfuge tubes. Cells were permeabilized with digitonin and the genomic DNA was cleaved and tagged by pA-Tn5 after incubation with histone H3 lysine 27 trimethyl primary antibody. Library was prepared using i5 and i7 universal primers and NEBNext High-fidelity 2x PCR Master mix. Before following the thermal cycling conditions recommended by the manufacturer, the samples were incubated at 72°C for gap filling.