ChIP-seq profiling of histone modifications (H3K4me3, H3K9ac, and H3K27me3) in seedling shoot tissues from 10 barley genotypes
Leaf samples were collected 4 hours after lights-on. Plants were grown in 35-well trays placed in growth chambers under a 16/8-hour light cycle at 20°C day/16°C night temperature for two weeks. 2 g of tissue was crosslinked with 1% formaldehyde for 15 min under vacuum using the Universal Plant ChIP-seq Kit (Diagenode). Nuclei were isolated, and chromatin was sheared. For ChIP, anti-H3K4me3 (ab213224, abcam; 04-745, Millipore; C15410003, Diagenode), anti-H3K9ac (ab4441, abcam), and anti-H3K27me3 (ab6002, abcam; C15410195, Diagenode) antibodies were used. Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB).