H3K27ac_RASH-1_ChIP-seq_Rep2
ChIP-seq were performed in RASH-1 cells. Cells were fixed in 1% formaldehyde for 10 min at room temperature and collected for nuclei extraction. Isolated nuclei were digested with MNase to obtain optimally sized chromatin fragments. Nuclear lysates were subjected to immunoprecipitation with antibody overnight. Magnetic beads were then incubated with the antibody-bound nuclear lysates for 2 hours at 4C. Beads were washed sequentially with ice-cold RIPA, RIPA500, LiCl, and TE buffers for 5 min each. DNA was eluted using elution buffer at 65C for 15 min. After reversal of cross-links, DNA was purified by phenol-chloroform-isoamyl alcohol extraction. Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit and sequenced on an Illumina NovaSeq platform (paired-end, 2  100 bp), with an average depth of 20 million reads per sample.