BstV2 I

Enzyme name BstV2 I
Prototype BbvII
Known analogs BbsI, BbvII, BpiI, BpuAI
SKU SE-E297
Turbo version Available
High-concentration version Not available
Restriction type Type IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GAAGAC(N)2▼ …3'
3'… CTTCTG(N)6▲ …5'
Source Bacillus stearothermophilus V2
Optimal buffer SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature 55 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
7575252510070
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA
Storage conditions 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 200 μg/ml BSA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.
Ligation After 5-fold overdigestion with enzyme more than 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 5 u.a. of enzyme for 16 hours at 55°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer Y, BSA
Notes High enzyme concentration may result in star activity. 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
  2. Unpublished data Sinichkina, S.A., Dedkov, V.S., Degtyarev, S.K Unpublished observations (2000).
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