[{"group": "General", "name": "Unit Definition One unit is defined as the amount of enzyme required to protect 1 µg of λ DNA in a total reaction volume of 20 μl in 1 hour at 37°C against cleavage by HaeIII restriction endonuclease. Reaction Conditions 1X GC Reaction Buffer Supplement with 160 µM S-adenosylmethionine (SAM) Incubate at 37°C 1X GC Reaction Buffer 50 mM NaCl 50 mM Tris-HCl 10 mM DTT (pH 8.5 @ 25°C) Control Protection Assay M.CviPI is incubated with 1 µg λ DNA in 20 μl 1X GC Reaction Buffer and 160 µM S-adenosylmethionine, for one hour at 37°C. The extent of protection by M.CviPI is determined by the addition of 30 μl NEBuffer 2 containing 10 units of HaeIII restriction endonuclease. Incubation for 1 hour at 37°C is followed by analysis on an agarose gel. Storage Buffer 15 mM Tris-HCl 0.2 M NaCl 1 mM DTT 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C Heat Inactivation 65°C for 20 minutes", "value": ""}]
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