Source: Acinetobacter calcoaceticus
Assayed on: Lambda DNA
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.
Optimal SE-buffer: Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
Activity in SEBuffers:
SEBuffer B 0 - 10%
SEBuffer G 0 - 10%
SEBuffer O 0 - 10%
SEBuffer W 0 - 10%
SEBuffer Y 100%
SEBuffer ROSE 80%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 0.05% Triton X-100; 50% glycerol;
Store at: -20°C.
Ligation: After 2-fold overdigestion with enzyme 90% of the DNA fragments can be ligated and 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.
Reagents supplied with enzyme: 10 X SE-buffer Y, BSA
Methylation sensitivity: Blocked by CG methylation
Inactivation: 20 minutes under 65°C
Notes: 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.