title: Illumina HiSeq 2500 sequencing; ChIP-seq of histone marks (H3K27me1, H3K27me3, H3K9me2) and of the H2A.W histone variant in Arabidopsis pol2a mutants.
design: ChIP-seq of histone marks (H3K27me1, H3K27me3, H3K9me2) and of the H2A.W histone variant in Arabidopsis pol2a mutants.
library_construction_protocol: Tissues were fixed in 1% (v/v) formaldehyde and flash frozen in liquid nitrogen. Plants were grown in vitro in a growth cabinet at 23°C, 50% humidity, using long day conditions (16h light, 8h dark). Tissues were homogenized in liquid nitrogen. After nuclei isolation and lysis, chromatin was sonicated in a Covaris S220 following manufacturer's instructions and shearing efficiency was verified on gel. Immunoprecipitation with specific antibodies was followed by reverse-crosslink and phenol-choloroform DNA purification. Libraries were constructed with the NuGEN Ovation Ultra Low System V2 kit, following the manufacturer's instructions.