DNA Methyltransferase M.BseCI
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DNA Methyltransferase M.BseCI
(concentration 5u/μl)

Technical Brochure



Description: M.ΒseCI modifies the N6 atom of the 3΄ adenine residue in the sequence 5΄-ATCGAT-3΄.


Source: M.BseCI is a methyltransferase purified from an E. coli strain that carries the BseCI methyltransferase gene (bseCIM) from Bacillus stearothermophilus, cloned in plasmid pBseCIM8 (1,2).


Reaction Buffer: 10 mM Tris-HCl (pH 7.4), 10 mM EDTA, 5 mM 2-mercaptoethanol, 0.02% Triton-X-100.

Unit definition: One unit is defined as the amount of enzyme required to protect 1μg of λ DNA in 1 hour at 55oC in a total reaction volume of 10μl against cleavage by BseCI restriction endonuclease.

Protection Assay Conditions: M.BseCI is incubated with 1μg of λ DNA in 10μl 1x M.BseCI buffer, supplemented with 80μM S-adenosylmethionine (SAM), for 1 hour at 55oC followed by 15 minutes at 70oC. The extent of protection by M.BseCI is determined by the addition of 40μl BseCI Reaction Buffer and 10 units of BseCI restriction endonuclease. Incubation at 55oC for 30 minutes is followed by analysis on agarose gel.

Note: M.BseCI exhibits 35% activity at 37oC.

Storage buffer: 50 mM Tris-HCl (pH 7.4), 10 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA and 50% glycerol. Store at -20oC.

Quality Control: Tested for the absence of endo- and exodeoxyribonucleases.


References: 1. Rina, M. and Bouriotis, V. (1993) Cloning, purification and characterization of the BseCI DNA methyltransferase from Bacillus stearothermophilus. Gene 133, 91-94.

2. Rina, M., Markaki, M. and Bouriotis, V. (1994) Sequence of the cloned bseCIM gene: M.BseCI reveals high homology to M.BanIII Gene 150, 71-73.


Reagents supplied: 10x M.BseCI buffer