NEB, M0215S, MspI Methyltransferase
Catalog Number: M0215S
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Pack Size5 units / pack
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| <b>Unit Definition</b> One unit is defined as the amount of enzyme required to protect 1 µg λ DNA in 1 hour at 37°C in a total reaction volume of 10 μl against cleavage by MspI restriction endonuclease. <b>Reaction Conditions</b> 1X MspI Methylase Reaction Buffer Supplement with 80 µM S-adenosylmethionine (SAM) Incubate at 37°C 1X MspI Methylase Reaction Buffer 50 mM Tris-HCl 100 mM NaCl 5 mM β-ME 10 mM EDTA (pH 7.5 @ 25°C) <b>Control Protection Assay</b> MspI Methyltransferase is incubated with 1 µg of λ DNA in 10 μl 1X MspI Methyltransferase Reaction Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection by MspI Methyltransferase is determined by the addition of 40 μl NEBuffer 2 supplemented with 10 mM MgCl 2 and 5 units of MspI restriction endonuclease. Incubation for MspI at 37°C is followed by analysis on agarose gels. <b>Storage Buffer</b> 50 mM Tris-HCl 50 mM NaCl 5 mM β-ME 10 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.5 @ 25°C <b>Heat Inactivation</b> No <b>Molecular Weight</b> Theoretical | 40397 daltons |
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