WGBS of Hordeum vulage subsp. Vulgare
Enzymatic methyl-seq tissues were homogenized in liquid nitrogen, and DNA was isolated with the DNeasy Plant Mini Kit (Qiagen). Libraries were prepared using the NEBNext Enzymatic Methyl-seq Kit (NEB) following the protocol for large DNA inserts. Therefore, 200ng genomic DNA was combined with 0.002 ng CpG methylated pUC19 DNA and 0.04 ng unmethylated lambda DNA. Fragmentation was done by using the Diagenode Bioruptor NGS in three rounds, 30s on, 90s off. Agilent Technologies 4200 Tape Station was used to determine the size distribution and concentration of the libraries.