|Source||Bacillus stearothermophillus T9|
|Description||N.Bst9I is a site specific DNA endonuclease that cleaves only one strand of double-stranded DNA substrate.|
|Unit definition||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.)|
|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.|
|Ligation||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|