High enzyme concentration results in star activity. To obtain 100% activity, SAM should be added to a final concentration 0.01 mM
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 | ↑(N)8AAGNNNNNCTT(N)13↑ |
Source | Flavobacterium aquatile Ob10 |
Assayed on | Lambda DNA |
Optimal buffer | W?(10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) +SAM |
Optimal temperature | 37°C |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
Ligations | After 3-fold overdigestion with enzyme 20% of the DNA fragments can be ligated and of these 80% can be recut. In the presence of 10%PEG ligation is better. |
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 | Methylation sensitivity not tested |
Thermal inactivation | enzyme is inactivated by incubation at 65°C for 20 mins. |
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