DMN-seq input of mES DNA site detection
Mouse embryonic stem cell (mESC) genomic DNA supplemented with 0.1% -DNA,  was fragmented using dsDNA Fragmentase, spiked with 200bp dsDNA oligos, and end-repaired/dA-tailed. After ligation with a customized DME adaptor, the DNA was digested with exonucleases to remove unligated or incorrectly processed fragments. The cleaned DNA was treated with recombinant DME enzyme under defined buffer conditions to excise 5mC. Resulting 5-phosphorylated fragments were ligated to a customized ssDNA adaptor using the TACS ligation method. Libraries were PCR-amplified, purified, and sequenced on the Illumina NextSeq 2000 platform.