Bst6 I

Enzyme nameBst6 I
PrototypeKsp632I
Known analogsEam1104I, EarI, Ksp632I
SKUSE-E239
Turbo versionNot available
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'… CTCTTCN▼ …3'
3'… GAGAAG(N)4▲ …5'
SourceBacillus stearothermophilus 6
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 temperature65 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
75755075100100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°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; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Storage at -20°C. Storage at -70°C is recommended for periods longer than 30 days..
LigationAfter 2-fold overdigestion with enzyme 80% of DNA fragments can be ligated. Of these 80% can be 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 65°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesTo obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  3. Unpublished data Abdurashitov, M.A., Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2000)
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