NEB, M0211S, EcoRI Methyltransferase
Catalog Number: M0211S
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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 EcoRI restriction endonuclease <b>Reaction Conditions</b> 1X rCutSmart™ Buffer Supplement with 80 µM S-adenosylmethionine (SAM) Incubate at 37°C 1X rCutSmart™ Buffer 50 mM Potassium Acetate 20 mM Tris-acetate 10 mM Magnesium Acetate 100 µg/ml Recombinant Albumin (pH 7.9 @ 25°C) <b>Control Protection Assay</b> rCutSmart Buffer is incubated with 1 µg of λ DNA in 10 μl 1X EcoRI Methyltransferase 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 EcoRI Methyltransferase is determined by the addition of 40 μl rCutSmart Buffer supplemented with 10 mM MgCl 2 and 5 units of EcoRI restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on an agarose gel. <b>Storage Buffer</b> 100 mM KPO4 200 mM NaCl 10 mM β-ME 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C <b>Heat Inactivation</b> 65°C for 20 minutes <b>Molecular Weight</b> Theoretical | 37911 daltons |
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