GpC Methylase (M.CviPI)
 
• For complete in vitro methylation of DNA for methylation analysis.
• Methylation of chromatin DNA for DNA accessibility studies.
• Inhibition of endonucleases with overlapping GpC sequence recognition.
• [3H]-labeling of DNA.
 
• Format: Provided in solution (4 units/µl) w/ 10X Reaction Buffer and 20X SAM
• Enzyme Commission Number: (EC 2.1.1.37)
• Storage: Store at -20°C for up to 12 months. Avoid repeated freeze/thawing of reagents. Prolonged storage is at ≤ -70°C.
• Unit Definition: One unit is defined as the amount of enzyme required to “protect” 1 µg of λ DNA against cleavage by HaeIII restriction endonuclease in a total reaction volume of 20 µl for 1 hour at 37°C.
• Reaction Conditions: GpC Methylase in 1X GpC Reaction Buffer w/ 600 µM SAM. Incubate reaction mixtures at 37°C.
• Inactivation: Heat-inactivate the enzyme at 65°C for 20 minutes.
• Standard Reaction Setup: The setup (below) is an example of a typical GpC methylase reaction in a 20 µl final reaction volume. The reaction volumes can be adjusted accordingly depending on experimental requirements (see protocol).
 
2 µl 10X GpC Reaction Buffer
1 µl 20X SAM (S-adenosylmethionine), 12 mM
4 µl DNA at 100-250 ng/µl
1 µl GpC Methylase (4 units/µl)
12 µl Water
  
Incubate at 37°C for 2 hours.
 

The GpC Methylase (EC 2.1.1.37)1 from Zymo Research completely methylates all cytosine residues (C5) in double-stranded, nonmethylated and hemi-methylated DNA having the dinucleotide sequence 5′…GpC…3′. The recombinant GpC Methylase (GST fusion) is isolated from an E. coli strain that expresses the methyltransferase gene from a Chlorella virus. The reaction conditions are optimized to maximize the processivity of the enzyme to ensure rapid, complete, and reproducible methylation of DNA for accurate DNA methylation analysis.

 

The GpC Methylase from Zymo Research catalyzes complete methylation of the GpC sites in DNA. Methylase activity of GpC Methylase from Zymo Research was tested for complete methylation of λ DNA using recommended reaction conditions. “Completion” of GpC methylation was assessed by resistance to digestion with a methylation sensitive endonuclease (HaeIII) and subsequently analyzed in an agarose gel. “M” is a 1kb DNA ladder (Zymo Research).

References:
1. Xu, M. et al. Nucleic Acids Res. 1998 Sept; 26(17):3961-3966

 
Product Catalog No. Size Price Store
GpC Methylase (M.CviPI) E2014 200 Units $
  E2015 1000 Units $
 
Components Available