BstNS I

Enzyme name BstNS I
Prototype NspI
Known analogs NspI, XceI
SKU SE-E251
Turbo version Available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… RCATG▼Y …3'
3'… Y▲GTACR …5'
Source Bacillus stearothermophilus NS
Optimal buffer SE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
1005010107550
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA
Storage conditions 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 10-fold overdigestion with enzyme >90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer B, BSA
Notes To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml Do not use BSA for long incubation.
References
  1. Unpublished data Abdurashitov, M.A., Bondar, T.S., Shevchenko, A.V., Degtyarev, S.K Unpublished observations (1996)
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