GSM8728324: Col-0 leaves, 3-4 week old, rep2, ChIP; Arabidopsis thaliana; ChIP-Seq
Leaves were fixed in 1% formaldehyde, 2 times for 15 min at room temperature in vacuum. Fixed tissue was ground to a fine powder in liquid nitrogen and nuclei were isolated. The crude nuclei were sonicated using Covaris S220 and centrifuged. The input sample was collected at this step. The rest of the supernatant was incubated with the GFP antibodies (#ab290, Abcam) followed by a capture with magnetic protein A beads (Dynabeads, Thermo) for immunoprecipitation. Bound chromatin was eluted from the beads. ChIP and Input samples were de-crosslinked and purified. The DNA sequencing libraries for both ChIP and Input samples were prepared using ThruPLEX DNA-Seq Kit (Takara Bio) and SMARTer DNA Unique Dual Index Kit (Takara Bio) following manufacturer's instructions. DNA double size selection 100-700 bp was performed using SPRIselect beads (Beckman Coulter).