GSM7881937: H3K27me3, Atsdr4l_5, 1 DAI, rep1; Arabidopsis thaliana; ChIP-Seq
Chromatins were fixed and extracted according to a published protocol (Song et al., 2016). Then, cross-linked DNA molecules with all proteins were fragmented by sonication using a Bioruptor Plus (Diagenode, Denville, NJ, USA) for 15 cycles, with each cycle consisting of 30 seconds ON and 90 seconds OFF at the HIGH setting. Sonicated chromatin was diluted by ChIP Dilution Buffer and then incubated overnight with HA antibody (CST, Danvers, MA, USA, 3724, 1:100 v/v dilution) and H3K27me3 antibody (Active Motif, Carlsbad, CA, USA, 61018, 1:100 v/v dilution) for precipitation of 3xHA-AtSDR4L-3xFLAG and H3K27me3 bound chromatin fragments, respectively. The captured chromatin was eluted in 200 μl Elution Buffer and then reserve crosslinked at 65°C for 6 h, followed by treatment of 100 μg/mL proteinase K at 55°C for 2 h before the phenol: chloroform: IAA (25:24:1 v/v, pH 8.0) extraction. DNA was precipitated and double‐size selected using 0.6 vol. and 1.3 vol. AMPure XP beads (Beckman Coulter, Brea, CA, USA, A63881) ChIP-seq libraries were constructed using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (NEB, Ipswich, MA, USA, E7645S)