| Recognition site | GAGTC(4/-)
|
| Source | Bacillus stearothermophillus T9 |
| Description | N.Bst9I is a site specific endonuclease that cleaves only one strand of T7 DNA on a double-strand DNA substrate. |
| Unit | One unit of the enzyme is the amount required to hydrolyze 1 μg
of T7 DNA in 1 hour at 55°C in a total reaction volume of 50μl. Concentrated enzymes are diluted to approximately 1000 units/ml with the buffer [10 mM Tris-HCl (pH 7.6); 50 mM KCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol] before determining their activity.
|
| Reaction buffer | SE-buffer N.Bst9I, (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 150 mM KCl; 1 mM DTT.) |
| Optimal temperature | 55oC |
| Storage conditions | 10 mM Tis-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C. |
| Ligations | After 5-fold overdigestion with N.Bst9 I, 90% of the T7 DNA fragments can be ligated and recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 2 units of N.Bst9I for 16 hours.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer N.Bst9I |
| Inactivation 20 minutes under | 80oC |
| Notes | High enzyme concentration and non optimal reaction conditions may result in star activity.
Incubation at 37°C results in 20% activity. |
| MSDS: | Download MSDS as PDF |
