GSM5237499: ZMP-HA-IP-br2 (B); Arabidopsis thaliana; ChIP-Seq
2 g of inflorescences was collected, ground to a fine powder in liquid nitrogen and cross-linked with 1% formaldehyde (Sigma Aldrich) for 20 min at room temperature with slow rotation. The chromatin was then fragmented to 300bp by sonication and the lysate was incubated with anti-HA polyclonal antibody (Sigma Aldrich, H6908) at 4 °C overnight. Subsequently, Dynabeads protein A and protein G were added followed by incubation for an additional 2 hr at 4 °C. The beads were washed five times for 5 min each at 4 °C and eluted twice by incubation in elution buffer (1% SDS, 0.1 M NaHCO3) at 65°C under rotation for 15 min each time. The cross-linking was reversed by incubation at 65 °C overnight, and the DNA was purified using a phenol:chloroform:isoamyl alcohol kit (Thermo Scientific, 17908) Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit (New England Biolabs) according to the manufacturer's protocol.