| 5'-A^C T A G T-3' |
|
| 3'-T G A T C^A-5' |
|
| Catalog # | B0808-05 |
| Concentration | 10u/ul |
| Source | Bacillus coagulans VS 29–022 |
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of AD2 DNA fragments in 1 hour at 37°C in 50ul of assay buffer. |
| 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 7.5 at 25°C), 300mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol. |
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug Ad2 DNA x 16 hours) with BcuI. |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with BcuI, more than 80% of the DNA fragments can be ligated at a 5'-termini concentration of 0.05uM. More than 90% of these can be recut. |
|
| 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 1.1% of white colonies were detected. |
|
| Stability during Prolonged Incubation | A minimum of 0.5units of enzyme is required for complete digestion of 1ug of Ad2 DNA in 16 hours at 37°C. |
|
| Thermal Inactivation | Enzyme is not inactivated by incubation at 80°C for 20min. |
|
| Digestion of Agarose-embedded DNA | A minimum of 10units of enzyme is required for digestion of 1ug of agarose-embedded Adenovirus-2 DNA in 16 hours. |
|
| Compatible Ends | Eco130I, NheI, XbaI, XmaJI |
|
| Number of Recognition Sites in DNA | |
|
| Ad2 | 3 |
| Lambda | 0 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 0 |
| pUC18/19 | 0 |
| pUC57 | 0 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 1 |
| pBluescriptIISK(-/+) | 1 |
| pACYC177 | 0 |
| pACYC184 | 0 |
