Source: Pseudomonas species-SE-G49
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: B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 100%
SEBuffer G 25 - 50%
SEBuffer O 10 - 25 %
SEBuffer W 25 - 50%
SEBuffer Y 75 - 100%
SEBuffer ROSE 40%
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); 200 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 50% glycerol;
Store at: -20°C.
Ligation: After 5-fold overdigestion with enzyme about 50% of the DNA fragments can be ligated and of these 95% can be 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 37°C.
Reagents supplied with enzyme: 10 X SE-buffer B
Methylation sensitivity: not tested
Inactivation: 20 minutes under 65°C
Notes: The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,25.