| 5'-G^G G C C C-3' |
|
| 3'-C C C G G^G-5' |
|
| Catalog # | B2945 |
| Unlike ApaI, Bsp120I produces DNA fragments with a 4-base 5'-extension. |
| Concentration | 10u/ul, 50u/ul(HC) |
| Source | Bacillus subtilis RFL120 |
| Buffer | 10mM Tris-HCl (pH 7.5), 10mM MgCl2 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. |
| Storage Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/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 Bsp120I. |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsp120I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.01uM. More than 95% 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 | PUC57 was digested at a unique site with 10units of enzyme for 16 hours. After religation and transformation 0.6% of white colonies were detected. |
|
| Methylation Effects | Bsp120I does not cut GGGCCm5C. Blocked by overlapping Dcm or 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. |
|
| Thermal Inactivation | Enzyme is inactivated by incubation at 80°C for 20min. |
|
| 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 | CfrI, Eco52I, NotI |
|
| Number of Recognition Sites in DNA | |
|
| Lambda | 1 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 0 |
| pUC18/19 | 0 |
| pUC57 | 1 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 1 |
| pBluescriptIISK(-/+) | 1 |
| pACYC177 | 0 |
| pACYC184 | 0 |
|
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-CpoI fragments in 1 hour at 37°C in 50ul of assay buffer. |
