title: GSM4600788: INPUT-Flag-H2B-set1-1; Saccharomyces cerevisiae; OTHER
library_construction_protocol: ChDIP assays were carried out as described previously (Nathan et al., 2006; Ryu et al., 2019). Preparation of chromatin fragments and elution steps were performed as described above for ChIP experiments. After overnight immunoprecipitation with anti-Flag agarose beads (Sigma, A2220) at 4°C, elution was performed with triple-Flag peptide (Sigma, F3290). While 5% of the elute was used as INPUT, the remaining 95% was incubated with anti-HA-conjugated agarose (Thermo Scientific, 26182) overnight at 4°C and then washed with FA lysis buffer four times. N-ethylmaleimide (NEM; Sigma, E3876) was added to buffers at a final concentration of 20 mM to prevent SUMO deconjugation. Approximately 5-10 ng of DNA is cleaned up using Ampure XP SPRI beads (Beckman Coulter Genomics). The DNA is then end-repaired, A-tailed, adapter ligated and PCR enriched (8-10 cycles) using the KAPA Hyper Library Preparation kit (KAPA Biosystems, Part#KK8504). Indexed libraries that meet appropriate cut-offs are quantified by qPCR using the Library Quantification Kit (KAPA Biosystems, Part#KK4854) and inserts size distribution by using the Caliper LabChip GX system. Samples with a yield of ≥0.5 ng/l are used for sequencing. ChDIP-seq
attributes.tag: GEO Accession
attributes.value: GSM4600788