BstV2 I

Enzyme nameBstV2 I
PrototypeBbvII
Known analogsBbsI, BbvII, BpiI, BpuAI
SKUSE-E297
Turbo versionAvailable
High-concentration versionNot available
Restriction typeType 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'
SourceBacillus stearothermophilus V2
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature55 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
7575252510070
Unit definitionOne 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 onLambda DNA
Storage conditions10 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.
LigationAfter 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo 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 sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesHigh 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 Unpublished data Sinichkina, S.A., Dedkov, V.S., Degtyarev, S.K Unpublished observations (2000).
// Код для некролога