GSM6559642: FL_H3K27Ac_rep2; Mus musculus; OTHER
CUT&Tag  was performed as previously described (55) with Hyperactive In-Situ ChIP Library Prep Kit for Illumina kit (Vazyme, TD901TD903) following the manufacturer's instructions. Briefly, a total of 50,000 cells were harvested, washed, and incubated with 10μl activated ConA beads per sample at room temperature for 10 min. Cells bound in beads were then incubated with primary antibodies (H3K27Ac, Abcam; IgG rabbit, Abcam) overnight at 4°C, followed by binding with secondary antibody at room temperature for 1hr. After washing twice with 800μl Dig-wash buffer, pA/G-Tn5 adapter complex was added to the sample in 100μl Dig-300 buffer to a final concentration of 1:250 and incubated on a nutator at room temperature for 1hr. Tagmentation was then performed at room temperature for 1hr and stopped with 10μl 0.5M EDTA, 3μl 10% SDS, and 2.5μl 20 mg/ml Proteinase K. Phenol-chloroform isoamyl alcohol extraction and ethanol precipitation of DNA was performed,Template DNA was extracted with DNA extraction beads,  followed by PCR amplification and Illumina sequencing.