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To obtain 100% activity, BSA should be added to the 1x 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 (dcm-, HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl. |
| Recognition sites | RG↑GWCCY |
| Source | Pseudomonas species PP |
| Assayed on | Lambda DNA (dcm-, HindIII-digest) |
| Optimal buffer | Y?(33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) |
| Optimal temperature | 37°C |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.|Store at -20°C.? |
| Ligations | After 5-fold overdigestion with enzyme more than 70% of the DNA fragments can be ligated and more than 80% of these can be recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 37°C. |
| Methlation effect | Blocked by overlapping Dcm methylation(CmCWGG):RGGWCCTGG. |
| Thermal inactivation | enzyme is inactivated by incubation at 65°C for 20 mins. |
| Campatible Ends | PpuMI, Psp5II |
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