GSM7290829: H4 control for H4K16ac ChIP-seq in ddm1 biol Rep2; Arabidopsis thaliana; ChIP-Seq
ChIP was performed as previously described (Parent et al., 2021), starting with 1g of seedlings. In brief, after crosslinking in 1% formaldehyde for 10 min, the tissue was ground to a fine powder in liquid nitrogen, chromatin was extracted with 1% SDS Tris-based lysis buffer and sonicated to ~200bp fragments. Chromatin was cleared with protein A magnetic beads and incubated overnight with the antibody listed below. Immune complexes were eluted with low, and then high salt buffers, before reversing crosslinks and purifying DNA fragments with ChIP DNA clean and concentrator kit (Zymo Research). Low input ChIP for MGH3-GFP was performed with ~10e8 pollen grains. Fixation was for 15 min and subsequent grinding with acid-washed glass beads. DNA fragments underwent a preliminary phenol-chloroform and ethanol precipitation step before clean up. NEBNext Ultra II DNA Library Prep Kit