title: GSM6722665: Input; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: 12-DAG seedlings were ground in liquid N2 to fine powder and resuspended in nuclei isolation buffer (60 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, pepstatin and complete protease inhibitors (Roche). Then, the samples were cross‐linked with 25 mM ethylene glycol bis succinimidyl succinate (EGS) by rotating for 20 min and with 1% formaldehyde by rotating for 10 min. The crosslinking of samples was stopped by 2M glycine for 10 min at room temperature. The chromatin was isolated and sheared into 200–500 bp fragments by sonication. For IP, the sonicated chromatin was incubated with 20 μl of anti‐GFP antibody (Thermo Fisher #A11122) overnight at 4°C while gentle rotating. BGISEQ platform Chip-seq library preparation:  1)End repair and adapter ligation:Fill the ends of IP/Input DNA followed by 5' phosphorylation. A 3' sticky-end with an overhanging A is formed, which could ligate with an overhanging T at the 3' end of the bubble adapter;  2)PCR amplification: PCR amplify the ligated products with specific primers;  3)Single-strand separation and circularization: Separate PCR products into single-strands by thermal denaturation. Then circularize the single-stranded DNA through a bridge primer to obtain single-stranded circular DNA library;  4)Sequencing: Amplified products DNA Nanoballs (DNBs) are used for sequencing.