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High enzyme concentration may result in star activity. To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml and SAM should be added to a final concentration 0.01 mM? 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 37°C in a total reaction volume of 50 μl. |
| Recognition sites | CTGAAG(N)16↑ |
| Source | E.coli strain that carries the cloned Acu I gene from Acinetobacter calcoaceticus SRW4 |
| 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 + SAM |
| Optimal temperature | 37°C |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
| Ligations | After 2-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated. Of these, 80% can be recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C. |
| Methlation effect | Methylation sensitivity not tested |
| Thermal inactivation | enzyme is inactivated by incubation at 65°C for 20 mins. |
| Campatible Ends | Eco57I |
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