title: GSM4222266: WT565_A; Saccharomyces cerevisiae; ChIP-Seq
library_construction_protocol: ChIP-seq was performed similarly as described (Rodriguez et al., 2014; PMID:  25271716 ) with following modifications. Zirconia/silica beads were used for bead beating. Chromatin was sonicated for three rounds of 15 min. S. pombe (strain 972h) chromatin used as a spike-in was prepared the same way as S. cerevisiae samples (including cross linking, bead beating and sonication) and kindly supplied by Toshi Tsukiyama. Each IP reaction contained 1 ug of S. cerevisiae chromatin, 10 ng of S. pombe chromatin and 20 ul of Protein G dynabeads (Invitrogen, #10004D) conjugated with 4 ul of anti-H3K18Ac antibody (EMD Milipore, #07-354) and the final volume was brought to 500 ul with FA buffer. 25 ul of the S. cerevisiae/S. pombe chromatin mix was transferred to a separate tube as an ?input? sample before addition of beads and combined with 25 ul of the stop buffer. Chromatin-antibody complexes were eluted with two 25 ul volumes of stop buffer. IP and input samples were processed together from this point. After Proteinase K digestion DNA was purified by phenol extraction (two step extraction for input samples) followed by ethanol precipitation in the presence of 20 ug glycogen. Pelleted DNA was washed with 70% ethanol and resuspended in 15 ul 10 mM Tris-HCl (pH 7.5). Sequencing libraries were prepared similarly as described (Warfield et al., 2017; PMID: 28918900) with few modifications. 2 ng of DNA was used as an input. Final adapter concentration during ligation was 6.5 nM. 15 cycles were used for library amplification.
attributes.tag: GEO Accession
attributes.value: GSM4222266