To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 50°C in a total reaction volume of 50 μl. |
Recognition sites | GCG↑C |
Source | Bacillus stearothermophilus HH |
Assayed on | Lambda DNA |
Optimal buffer | Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA |
Optimal temperature | 50°C |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol; Store at 20°C |
Ligations | After 40-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut |
Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of DNA with 100 u.a. of enzyme for 16 hours at 65°C. |
Methlation effect | Blocked by 5`-G(5mC)GC-3`/3-CG(5mC)G-5` or 5`-G(5mC)GC-3`/3`-CGCG-5` methylation.|Not blocked by 5`-GCG(5mC)-3`/3`-(5mC)GCG-5` or 5`-GCG(5mC)-3`/3`-CGCG-5` methylation. |
Thermal inactivation | NO |
Campatible Ends | AspLEI , CfoI, GlaI^, HhaI, R9529^, HinP1I^, HspAI?^ |
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