10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol.
Incubate at 37°C.
1XSEBuffer Y (pH 7.9 @ 25ºC):
33 mM Tris-Ac, 66 mM KAc, 10 mM MgAc,1 mM DTT
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
Enzyme is active in presence of EDTA.
Quality Control Assays
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.
No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 0.5 units of enzyme for 3
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.
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