NOMe-Seq analysis of fixed budding yeast nuclei with two DNA methyltransferases

NOMe-Seq analysis was performed with two cytosine C-5 DNA methyltransferases recognizing GC and CC dinucleotides Spheroplasts were prepared with treating S288C cells with Zymolyase 100T. Then, 1% formaldehyde was added to fix the cells. After washing the cells with 1 M sorbitol, the yeast nuclei were prepared with a solution containing 0.1% NP-40. The nuclei were suspended in methylation buffer, and cytosine C-5 DNA methyltransferase was added. The solution was incubated at 37°C for 1 hr and collected nuclei. Then genomic DNA was extracted, and library preparation was performed with tPBAT protocol (Miura F et al., NAR, 2019)