| 5'-A T^C G A T-3' |
|
| 3'-T A G C^T A-5' |
|
| Catalog # | B3050 |
| Concentration | 10u/ul |
| Source | Bacillus subtilis 15 |
| Buffer | 33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C |
| Diluent Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used. |
| Storage Buffer | 10mM Tris-HCl (pH 8.0 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol. |
| 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. |
|
| 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. |
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| 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. |
|
| 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. |
|
| Methylation Effects | Bsu15I cuts neither ATCGm6AT nor ATm5CGAT. Blocked by overlapping Dam methylation. Blocked by CG methylation. |
|
| 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°C. |
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| Thermal Inactivation | Enzyme is inactivated by incubation at 65°C for 20min. |
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| 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 |
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| Number of Recognition Sites in DNA | |
|
| Lambda | 15 |
| PhiX174 | 0 |
| M13mp18/19 | 2 |
| pBR322 | 1 |
| pUC18/19 | 0 |
| pUC57 | 0 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 1 |
| pBluescriptIISK(-/+) | 1 |
| pACYC177 | 1 |
| pACYC184 | 1 |
|
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 37°C in 50ul of assay buffer. |
