GSM6912125: Yeast_HPRT1R_integrated_IgG_ChIP_rep2; Saccharomyces cerevisiae; ChIP-Seq
Cells were pelleted at 3,000 rcf for 5 minutes, washed twice with water, and resuspended in spheroplasting buffer (1.4 M Sorbitol, 40 mM HEPES-KOH pH 7.5, 0.5 mM MgCl2, 0.5 mM 2-mercaptoethanol). Spheroplasting was performed by adding 0.125 U/μL Zymolyase (Zymo Research E1004) and incubating at 37oC for 45 minutes on a rotator. Nuclei were prepared as previously described (Orsi, G.A., et al., Curr Protoc Mol Biol, 2015). Resuspended nuclei were split into aliquots of ~108 nuclei each and snap frozen in liquid nitrogen. ~10^8 frozen nuclei were thawed and processed for CUT&RUN using the CUTANA™ ChIC/CUT&RUN kit (EpiCy-pher 14-1048) according to the manufacturer's protocol. Antibodies were  used at 0.5 μg: rabbit IgG negative control (EpiCypher 13-0042), H3K4me3 (EpiCypher 13-0041). Sequencing libraries were prepared using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (New England Biolabs E7645L) according to the manufacturer's protocol.