title: GSM4600857: IP-Set3-TAP-WT-1; Saccharomyces cerevisiae; ChIP-Seq
library_construction_protocol: ChIP experiments were performed as described previously (Ryu & Ahn, 2014; Ryu et al, 2016). TAP-tagged Set3 proteins were precipitated with IgG-Sepharose beads (GE Healthcare, 17-0969-01). Formaldehyde was added to a final concentration of 1% for 20 min. Cross-linking was quenched by addition of glycine to 240 mM. Cells were collected by centrifugation, washed in 1× TBS, and lysed with glass beads in FA lysis buffer (50 mM HEPES-KOH at pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% Na deoxycholate, 0.1% SDS, 1 mM PMSF). Chromatin was sheared by sonication, therefore the average fragment size was between 200 and 700 bp. 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.
attributes.tag: GEO Accession
attributes.value: GSM4600857