Source: Bacillus stearothermophilus DS
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 65°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.)
Activity in SEBuffers:
SEBuffer B 0 - 10%
SEBuffer G 75 -100%
SEBuffer O 50 - 75%
SEBuffer W 25 - 50%
SEBuffer Y 100%
SEBuffer ROSE 100%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 65°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Store: at -20°C.
Ligation: After 10-fold overdigestion with enzyme 95% 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 20 u.a. of enzyme for 16 hours at 65°C.
Reagents supplied with Enzyme: 10 X SE-buffer Y
Methylation sensitivity: not tested
Inactivation: 20 minutes under 80°C
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