Source: Bacillus stearothermophilus P
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 50°C in a total reaction volume of 50 μl.
Optimal SE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 50 - 75%
SEBuffer G 100%
SEBuffer O 75 - 100%
SEBuffer W 50 - 75%
SEBuffer Y 50 - 75%
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: 50°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 10 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol;
Store: at -20°C.
Ligation: After 5-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 10 u.a. of enzyme for 16 hours at 50°C.
Reagents Supplied with Enzyme: 10 X SE-buffer G
Methylation sensitivity: Blocked by CG methylation.
Inactivation: 20 minutes under 65°C