Source: Bacillus stearothermophilus V1
Assayed on: pBR322 DNA
Unit definition: One unit of the enzyme is the amount required to hydrolyze 1 μg of pBR322 DNA in 1 hour at 55°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 75 -100%
SEBuffer G 100%
SEBuffer O 50-100%
SEBuffer W 75 -100%
SEBuffer Y 75-100%
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: 55°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol.
Store: at -20°C.
Ligation: After 3-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of DNA with 2 u.a. of enzyme for 16 hours at 55°C.
Reagents supplied with Enzyme: 10 X SE-buffer G
Methylation sensitivity: not tested
Inactivation: 20 minutes under 80°C
Notes: at 37°C activity is 10% from maximum.
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