In vitro methylation of DNA for methylation analysis.
Inhibition of endonucleases with overlapping CpG sequence recognition.
[3H]-labeling of DNA.
Description
CpG Methyltransferase (M.SssI) is one of the basic tools used in epigenetic studies. The enzyme methylates the 5’ 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 restriction times without compromising the reaction efficiency. The enzyme completes modification of all CpGs in 15 min 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.
Storage buffer
CpG Methyltransferase (M.SssI) is supplied in: 10 mM potassium phosphate (pH 7.0 at 25°C), 400 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.
Formulation
One µl of M.SssI protects 1 µg of genomic DNA from digestion with HpaII in 15 min at 37°C in 50 µl of recommended reaction buffer.
Quality Control
Prolonged incubation No detectable change in DNA fragment band pattern observed following agarose gel electrophoresis of SmaI linearized pUC19 DNA incubated for 16 hours with 1 µl of M.SssI.
Labeled Oligonucleotide (LO) Assay No detectable degradation of single-stranded or double-stranded labeled oligonucleotides observed after incubation with 1 µl of M.SssI for 4 hours at 37°C, analysis by polyacrylamide gel electrophoresis and phosphoimaging