Bsu I

Enzyme nameBsu I
PrototypeBciVI
Known analogsBciVI
SKUSE-E581
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'… GTATCC(N)6▼ …3'
3'… CATAGG(N)5▲ …5'
SourceBacillus sphaericus
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 temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
7550102510010
Unit definitionOne 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 onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 0.05% Triton X-100, 50% glycerol; Store at -20°C.
LigationAfter 5-fold overdigestion with enzyme ~10% of DNA fragments can be ligated with T4 DNA Ligase and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 4 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer Y
NotesWhen using a buffer other than the optimal (suppied) SEBuffer, it may benecessary to add more enzyme to achieve complete digestion.
References

No publications found.

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