Bmu I

Supplied in:

10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol.

Reaction Conditions:

1XSEBuffer Y

Incubate at 37°C.

1XSEBuffer Y  (pH 7.9 @ 25ºC):

33 mM Tris-Ac, 66 mM KAc, 10 mM MgAc,1 mM DTT

Unit Definition:

One unit is defined as the amount of  enzyme required to digest 1 µg of l DNA /Hind III in 1 hour at 37°C in a total reaction volume of  50µl. 

Note:

Enzyme is active in presence of EDTA.

Quality Control Assays

Ligation:

After 2-fold overdigestion with Bmu I ~75% of the λ DNA fragments can be ligated and  95% of these can be recut. Overnight digest with BmuI is not recommended. A 50 µl reaction containing 1 µg of 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.

Oligonucleotide Assay:

No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 0.5 units of enzyme for 3 hours.

Enzyme Properties

Activity in SEBuffers:

SEBuffer B    75-100%

SEBuffer G    75--100%

SEBuffer O    25-50%

SEBuffer W   10-25%

SEBuffer Y       100%

SEBuffer ROSE  25% 

When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

Heat Inactivation:

Yes   (65°C for 20 minutes)

Reagents supplied with enzyme:

10X SEBuffer Y