Dilution Buffer
10mM Tris-HCl (pH 7.4 at 25 degree C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods-the Storage Buffer should be used.
Digestion of Agarose-embedded DNA
A minimum of 5units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible Ends
AciI, Bsp119I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, TaqI, XmiI
Methylation Effects
Bsu15I cuts neither ATCGm6AT nor ATm5CGAT. Blocked by overlapping Dam methylation. Blocked by CG methylation.
Blue/White Cloning Assay
The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.5% of white colonies were detected.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsu15I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.15uM. More than 95% of these can be recut.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 240-fold overdigestion (15u/ug lambda DNA x 16 hours) with Bsu15I.
Stability During Prolonged Incubation
A minimum of 0.1units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37 degree C.
Storage Buffer
10mM Tris-HCl (pH 8.0 at 25 degree C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol.
Thermal Inactivation
Enzyme is inactivated by incubation at 65 degree C for 20min.
Unit Definition
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 37 degree C in 50ul of assay buffer.
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