title: GSM7809260: H3-NHS-r1; Arabidopsis thaliana; ChIP-Seq
library_name: GSM7809260
library_construction_protocol: Seedlings were crosslinked under vacuum in PBS buffer with 1 % (w/v) formaldehyde for 10 min on ice. Chromatin was extracted as described previously (Kaufmann et al., 2010)⁠. Chromatin was fragmented using a Bioruptor (Diagenode – 20 cycles 30sec on/30sec off). For single ChIP, chromatin was immunoprecipitated overnight at 4°C with IgG (ThermoFisher 026102), anti-panH3 (Diagenode C15200011) or anti-GFP (Chromotek PABG1) antibodies. Antibody-chromatin complexes were purified with Dynabeads (ThermoFischer 10002D) and successively washed with low salt (150 mM NaCl, 1 % (v/v) Triton X-100), high salt (500 mM NaCl, 1 % (v/v) Triton X-100), LiCl buffer (250 mM LiCl, 1 % (v/v) NP-40) and TE buffer. Chromatin was eluted with single ChIP elution buffer (1 % (w/v) SDS, 0.1 M NaHCO3). ChIP DNA was generated and libraries were prepared using NEBNext Ultra II FS DNA Library preparation kit (NEB E7645). Quality control was performed with TapeStation D1000 ScreenTape system (Agilent) and a Qubit fluorometer (Thermo Fisher).