Home | Molecular Biology | Molecular Biology Applications | Epigenetics | Epigenetic Enzymes

CpG Methyltransferase (M.SssI)


CpG Methyltransferase (M.SssI) is one of the basic tools used in epigenetic studies.
  
Loading...

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.

Highlights

  • High efficiency – complete in vitro methylation of all CpGs in non-methylated and hemi-methylated DNA
  • Fast – reaction is completed in 15 minutes at 37°C
  • Stable – at -20°C for 2 years

Applications

  • Epigenetics studies
  • In vitro methylation of DNA for methylation analysis
  • Inhibition of endonucleases with overlapping CpG sequence recognition.
  • [3H]-labeling of DNA
  
Storage Condition-20 C
HazardousNo
FormulationOne 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 ControlProlonged 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.
Storage BufferCpG 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.
Plasmid DNA methylated for 15 min using Thermo Scientific M.SssI is protected from HpaII cleavage.

Plasmid DNA methylated for 15 min using Thermo Scientific M.SssI is protected from HpaII cleavage.

Plasmid DNA methylated for 15 min using Thermo Scientific M.SssI is protected from HpaII cleavage.

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 HpaII
3 – after digestion with MspI


Protocols and Product Manuals

pdf   CpG Methyltransferase (M.SssI) - Product Information