GSM8287122: WT_Vehicle_H3K9ac_rep1; Saccharomyces cerevisiae; ChIP-Seq
H3, H3K9ac, and RNAPII ChIP-seq was performed as described previously (Ribaud et al., 2012; PMID: 21952045) with the following modification. A defined amount (5% of total chromatin) of previously cross-linked chromatin from S.pombe was added to S.cerevisiae chromatin prior to immunoprecipitation step. 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)