GSM8287152: Stb3-ChIP_WT_Control; Saccharomyces cerevisiae; ChIP-Seq
ChIP protocol was performed as previously described (Zencir et al., 2020, Ribaud et al., 2012). The cells were crosslinked with 1% formaldehyde for 10 min and quenched by adding 125 mM glycine for 5 min at room temperature. Briefly, crosslinked cells were lysed by bead beating, chromatin was sonicated, and the soluble fraction was incubated with the appropriate antibodies for 1 hr at 4°C. Magnetic beads coupled to IgG against rabbit or mouse (Invitrogen, Dynabeads™ M-280 Sheep Anti-Rabbit or Anti-Mouse IgG) were added into the samples and incubated for additional 3 h at 4°C. A defined amount (5% of total chromatin) of crosslinked and sonicated chromatin from S. pombe was added to S. cerevisiae chromatin prior to the immunoprecipitation step. ChIP was performed using the following antibodies: anti-histone H3 ChIP, (Abcam ab1791); anti-H3K9ac (Millipore 07-352); anti-RNAPII (Abcam ab5131); fanti-Flag (Sigma F3165); anti-Myc (a homemade antibody, clone 9E10). The DNAs were purified using the MinElute PCR Purification Kit (Qiagen) before library preparation. TruSeq ChIP library preparation kit (Illumina)