GSM8493713: CKO H3K27ac CUT&Tag Rep1; Mus musculus; OTHER
DNA was extracted by DNA clean beads CUT&Tag sample preparation and experiments were performed using the Hyperactive In-Situ ChIP Library Prep Kit for Illumina (Vazyme, TD901-01) as described before (Kaya-Okur et al., 2019) with minimal changes. Briefly, cells were immobilized using ConA beads. Then, cells were permeabilized and incubated with different primary antibodies (anti-H3K27ac, Abcam 177178) overnight for 4°C and then treated with secondary antibodies (Goat anti-Rabbit IgG) for 1 h at room temperature. Afterward, cells were incubated with pG-Tn5 transposase for 1 h and then cells were washed twice and incubated at 37 °C for 1 h. DNA of each sample was extracted using phenol-chloroform-isoamyl alcohol and amplified by PCR. All CUT&Tag libraries were sequenced on the Illumina Nova Seq 6000 platform at PE150 mode.