title: GSM8584182: WT,24h,ChIP-seq; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: Total 4 g of 18-d-old plants was used for each sample. Add c. 2 g of ground tissue to a 50 ml Falcon tube containing 25 ml nuclear isolation buffer (10 mM HEPES pH 8.0, 1 M sucrose, 5 mM KCl, 5 mM MgCl2, 5 mM EDTA, 0.6% Triton X-100, 0.4 mM PMSF, and 1 × Mini-Complete cocktail). Then, crosslinked with 1% formaldehyde for 20 min and used for chromatin extraction. After 15 min sonication (45 s on/15 s off, at high energy level) by Covaris S220 (Thermo Fisher, Shanghai, China) and centrifugation for 15 min at 12 000 g, the supernatant was incubated with 5 μg anti-H4K16ac antibody (PTM-122; PTM Bio, Hangzhou, China). Nipponbare rice nucleosomes were added as spike-in with 1 : 50 ratio. After one night, the supernatant was added 40 μl Protein A/G beads (SA032005; Smart Lifesciences, Changzhou, China) and incubated 2 h with rotation at 4°C. After extensive washing, immunoprecipitated chromatin was reverse-crosslinked and purified for sequencing. For ChIP-Seq, DNA from chromatin immunoprecipitation was used to construct sequencing libraries following the protocol provided with Scale ssDNA-Seq Lib Prep Kit for Illumina V2 (RK20228; Abclonal) and sequenced on an Illumina NovaSeq 6000 instrument (Annoroad).
identifiers.namespace: GEO
identifiers.id: GSM8584182_r1