GSM8728250: HB34_IP_1; Arabidopsis thaliana; ChIP-Seq
Briefly, about 0.5-g 10-d-old seedlings were crosslinked with 1% formaldehyde and stopped with 125 mM glycine. Samples were ground and resuspended in the nuclei isolating buffer (10 mM Tris pH 8.0, 10 mM Sodium butyrate, 400 mM Sucrose, 0.1 mM PMSF, 5 mM β-mercaptoethanol, 1×cocktail protein inhibitor freshly). The isolated nuclei were resuspended with 1 mL lysis buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 1 mM EDTA, 10 mM Sodium butyrate, 0.1% deoxycholate, 0.1% SDS, 1% Triton X-100, 1 mM PMSF and 1×protease inhibitor cocktail freshly) and sonicated into 200-500 bp fragments by Diagenode Bioruptor (high setting, 18 cycles，30s on and 30s off). Subsequently, the sonicated chromatin was incubated with GFP-antibodies (Abcam, ab290) overnight at 4°C. The recovered DNAs in ChIP assay were used to library construction with the Scale ssDNA-seq Lib Prep Kit (ABclonal, RK20228). High-throughput sequencing was carried out using Illumina Hiseq-PE150 by Novogene Corporation (Beijing, China).