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The enzyme methylates the C5 position on the base moiety of all cytosine nucleotides contained in unmethylated or hemimethylated double stranded DNA in a 5’-CpG-3’ context.
Thermo Scientific M.SssI is specifically formulated for fast reaction times without compromising the reaction efficiency. The enzyme completes modification of all CpGs in 15 minutes at 37°C. In addition, the enzyme has been specifically validated for use with genomic DNA – the primary substrate in epigenetic studies.
M.SssI is supplied with an optimized 10X reaction buffer and 50X S-adenosylmethionine (SAM) as a cofactor.
0.5 µg of linearized plasmid DNA (pUC19/SmaI) were methylated using M.SssI from different suppliers according to vendor recommendations. The methylated plasmid DNA was mixed with 0.5 µg of human blood genomic DNA and digested in parallel with methylation-sensitive HpaII restriction enzyme and methylation insensitive isoschizomer MspI. Methylated plasmid DNA and naturally methylated gDNA were protected from cleavage with HpaII and cleaved with insensitive MspI.1 – undigested DNA (control)2 – after digestion with HpaII3 – after digestion with MspI