GSM5849600: HID1_ChIRP_Rep1; Arabidopsis thaliana; OTHER
GSM5849600
0.3 g seeds were used for each assay. The materials were fixed by two methods in the dark room. For reversible chemical cross-linking, fixing was performed with 1% (v/v) formaldehyde in PBS at room temperature for 20 min under vacuum. For non-reversible cross-linking, fixing was performed with 1% (v/v) glutaraldehyde in PBS at room temperature for 20 min under vacuum. For both cross-linking methods, the reactions were quenched with glycine (0.125 M final concentration) for 5 min under vacuum. Antisense DNA probes were designed against the full-length HID1 sequence and biotinylated at the 3′ end (Invitrogen). Unlabeled probes were used as the control. DNA was purified with the ChIP DNA Clean & Concentrator kit (D5205, Zymo research) according to the manufacturer's instructions, and the purified DNA was subjected to high-throughput sequencing. All ChIRP-seq libraries were generated using the NEBNext® Ultra II DNA Library Prep Kit for Illumina (E7645S, NEB) according to the manufacturer's instructions. All libraries were sequenced on a HiSeq 2500 System with paired-end 150 bp reads.