title: GSM5106475: REF6-HA-24h_rep1; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: For REF6-HA binding ChIP, chromatin was resuspended with ChIP lysis buffer (50mM HEPES (pH7.5), 150mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% deoxycholate, 0.5% SDS, and 1×protease inhibitor cocktail (Sigma, S8830)) and sheared to generate 200-500bp fragments with Bioruptor (Diagenode) for 30 cycles (30s/on, 30s/off) at high power. Sheared chromatin was diluted four times with ChIP dilution buffer (50mM HEPES (pH7.5), 150mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% deoxycholate, 0.01% SDS, and 1×protease inhibitor cocktail (Sigma, S8830)) and subjected to immunoprecipitation with anti-HA antibody (CST, 3724). After antibody incubation, Protein A Dynabeads (Life technologies, 10002D) were added to collect the immunocomplexes, followed by elution, reverse cross-linking and DNA purification. Library was prepared with VAHTS universal DNA library prep kit for illumina (Vazyme, ND607) according to the manufacture's instruction