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SibEnzyme Ltd. :: Display product information: N.Bst9 I
Name
N.Bst9 I
Catalog numberE401E402
Package, u.a.100500
Concentration, u.a./ml2000-50002000-5000

Recognition siteGAGTC(4/-)
DescriptionN.Bst9I is a site specific endonuclease that cleaves only one strand of T7 DNA on a double-strand DNA substrate.
OriginBacillus stearothermophillus T9
Reaction bufferSE-buffer N.Bst9I, (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 150 mM KCl; 1 mM DTT.)
Storage conditions10 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 -20C.
LigationAfter 5-fold overdigestion with N.Bst9 I, 90% of the T7 DNA fragments can be ligated and recut.
Non-specific activityNo nonspecific activity was detected after incubation of 1 μg of T7 DNA with 2 units of N.Bst9I for 16 hours.
Optimum temperature55 oC
UnitOne unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 55C in a total reaction volume of 50ul. 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.
Inactivation 20 minutes80oC

Note: High enzyme concentration and non optimal reaction conditions may result in star activity.
Incubation at 37C results in 20% activity.

References:

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