Bmu I

Enzyme name Bmu I
Prototype BfiI
Known analogs BfiI, BmrI
SKU SE-E487
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'… ACTGGG(N)5▼ …3'
3'… TGACCC(N)4▲ …5'
Source Bacillus megaterium S87
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 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
7575251010025
Unit definition One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on Lambda DNA (HindIII-digest)
Storage conditions 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C
Ligation After 2-fold overdigestion with enzyme about 75% of the DNA fragments can be ligated and 95% may be recut.
Nonspecific hydrolysis Overnight digest with BmuI is not recommended. A 50 μl reaction containing 1 μg of Lambda DNA and 0.5 units of enzyme incubated for 4 hours resulted in the same pattern of DNA bands as a reaction incubated for 1 hour.
Methylation sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer Y
Notes Enzyme is active in presence of EDTA.
References

No publications found.
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