title: GSM9165851: Arabidopsis thaliana, Col, AR seed, H3K27me3 ChIP; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: Seed chromatin was isolated for native-ChIP as previously described by Müller et al. (2012) with the following modifications. Whole seeds rather than isolated embryos were used for chromatin extraction. Chromatin was fragmented using MNase (NEB, MS247S) by incubating at 37°C for 20 minutes (min), extending the incubation time from the 8 min described in the original protocol. For immunoprecipitation of histone protein-DNA complexes, the following antibodies were used: α-H3K4me3 (Millipore, 07-473), α-H3K9me3 (Millipore, 07-442), and α-H3K27me3 (Millipore, 07-449). ChIPed DNAs were purified by the MinElute PCR Purification Kit (Qiagen, 28006), followed by Proteinase K (Roche, 03115828001) treatment. ChIP-seq libraries were constructed using the TruSeq Library Preparation Kit (Illumina, IP-202-1012) and sequenced on the Illumina HiSeq4000 platform by Macrogen (Seoul, South Korea).