| Catalog # | B2935 |
| Sequence | 5'-T C G^C G A-3' |
| 3'-A G C^G C T-5' |
| Source | Bacillus megaterium RFL68 |
| Concentration | 10u/ul |
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. |
| Diluent Buffer | 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. |
| Storage Buffer | 10mM Tris-HCl pH 7.5, 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol. |
| R1625 Restriction Enzyme Buffer A, 10X | 1X buffer formulation - 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. |
| R1625-03 Restriction Enzyme Buffer D, 10X | 1X buffer formulation - 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM sodium chloride and 0.1mg/ml BSA. |
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 15-fold overdigestion (15u/ug lambda DNA x 1 hour) with Bsp68I (see Star Activity). |
|
| Ligation/Recutting Assay | |
| After 10-fold overdigestion (0.6u/ug DNA x 17 hours) with Bsp68I, more than 80% of the digested PhiX174 DNA fragments can be ligated in a reaction mixture containing 20-40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.17uM. 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 10 units of Bsp68I for 4 hours. |
|
| Blue/White Cloning Assay | |
| pUC57 was digested at a unique site with 10 units of Bsp68I for 1 hour. After religation and transformation < 1% of white colonies were detected. |
|
| Star Activity | |
| An excess of Bsp68I (20u/ug DNA x 1 hour) may result in star activity. |
|
| Methylation Effect | |
| Dam | May overlap - no effect |
| Dcm, EcoKI, EcoBI | Never overlaps - no effect |
| CpG | Completely overlaps - blocked |
|
| Incubation Temperature | |
| Incubate at 37°C. |
|
| Stability during Prolonged Incubation | |
| A minimum of 0.2 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
|
| Digestion of Agarose-embedded DNA | |
| A minimum of 5 units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Thermal Inactivation | |
| Inactivated by incubation at 65°C for 20 min. |
|
| Number of Recognition Sites in DNA | |
| Lambda | 5 |
| PhiX174 | 2 |
| M13mp18/19 | 0 |
| pBR322 | 1 |
| pUC18/19 | 0 |
| pUC57 | 1 |
| pTZ19R/U | 0 |
