Bst6 I

Enzyme name Bst6 I
Prototype Ksp632I
Known analogs Eam1104I, EarI, Ksp632I
SKU SE-E239
Turbo version Not 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'… CTCTTCN▼ …3'
3'… GAGAAG(N)4▲ …5'
Source Bacillus stearothermophilus 6
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 65 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
75755075100100
Unit definition One 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 on Lambda DNA
Storage conditions 10 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..
Ligation After 2-fold overdigestion with enzyme 80% of DNA fragments can be ligated. Of these 80% can be 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 65°C.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer Y, BSA
Notes To 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 Unpublished data Abdurashitov, M.A., Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2000)
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