NEB, M0224S, HaeIII Methyltransferase

[{"group": "General", "name": "Unit Definition 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 HaeIII restriction endonuclease. Reaction Conditions 1X HaeIII Methyltransferase Reaction Buffer Supplement with 80 µM S-adenosylmethionine (SAM) Incubate at 37°C 1X HaeIII Methyltransferase Reaction Buffer 50 mM Tris-HCl 50 mM NaCl 10 mM DTT (pH 8.5 @ 25°C) Control Protection Assay HaeIII Methyltransferase is incubated with 1 µg of λ DNA in 10 μl of 1X HaeIII 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 is determined by the addition of 40 μl NEBuffer 2.1 and 10 units of HaeIII restriction endonuclease. Incubation for 1 hour at 37°C is followed by analysis on an agarose gel. Storage Buffer 50 mM Tris-HCl 1 mM DTT 10 mM EDTA 200 µg/ml BSA 50% Glycerol 50 mM KCl pH 7.4 @ 25°C Heat Inactivation 65°C for 20 minutes", "value": ""}]