10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 200 mg/ml BSA;
2-mercaptoethanol; 50% glycerol.
1XSEBuffer G. Incubate at 37°C.
G (pH 7.6 @ 25ºC)
10 mM Tris-HCl 50 mM NaCl
10 mM MgCl2 1 mM DTT
The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA .
Kro I doesn`t cleave DNA modified with MspI DNA-methyltransferase.
Unit Definition: One unit is defined as the amount of enzyme required to hydrolyze completely1 μg of linearized plasmid pMHpaII 1 in
1 hour at 37°C in a total reaction volume of 50 μl.
DNA pMHpaII 1/DriI is a linearized plasmid pMHpaII 1. pMHpaII 1 carries a gene of DNA-methyltransferase M.HpaII, which methylates sites
5`-CCGG-3` producing 5`-C(5mC)GG-3`/3`-GG(5mC)C-5`, and includes three canonical sites 5`-GC(5mC)GGC-3`/3`-CGG(5mC)CG-5`.
Reagents Supplied with Enzyme: 10XSEBuffer G
Heat Inactivation: Yes (65°C for 20 minutes)