NEB, M0222S, dam Methyltransferase

[{"group": "General", "name": "Unit Definition One unit is defined as the amount of enzyme required to protect 1 µg ( dam -) Lambda DNA in 1 hour at 37°C in a total reaction volume of 10 μl against cleavage by MboI restriction endonuclease. Reaction Conditions 1X dam Methyltransferase Reaction Buffer Supplement with 80 µM S-adenosylmethionine (SAM) Incubate at 37°C 1X dam Methyltransferase Reaction Buffer 50 mM Tris-HCl 5 mM β-ME 10 mM EDTA (pH 7.5 @ 25°C) Control Protection Assay dam Methyltransferase is incubated with 1 µg of ( dam- ) Lambda DNA in 10 μl of 1X dam Methyltransferase Buffer, supplemented with 80 µM S-adenosylmethionine, for 1 hour at 37°C followed by 15 minutes at 65°C. The extent of protection is determined by addition of 40 μl 1X NEBuffer 3 supplemented with 10 mM MgCl 2 and 10 units of MboI restriction endonuclease. Incubation at 37°C for 1 hour is followed by analysis on an agarose gel. Storage Buffer 50 mM Tris-HCl 50 mM KCl 1 mM DTT 10 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.5 @ 25°C Heat Inactivation 65°C for 20 minutes", "value": ""}]