| 5'-G C^T N A G C-3' |
|
| 3'-C G A N T^C G-5' |
|
| Catalog # | B2650 |
| Source | Bacillus pumilus RFL1102 |
| 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. |
| Concentration | 10 units/ul |
| R1625 Restriction Enzyme Buffer A, 10X, | 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. |
| Double Digests | R1625 Restriction Enzyme Buffer A, 10X (Catalog No: R1625) is provided to simplify buffer selection for double digests. Y+/Tangle Buffer is supplied as a liquid in 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37°C. |
| |
| Dilution/Storage Buffer:: 10mM Tris-HCl, pH 7.4 at 25°C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Quality Control | Overdigestion Assay: |
| No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Bpu1102I. |
| |
| Ligation/Recutting Assay: |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bpu1102I, more than 80% of the DNA fragments can be ligated in 20-40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.2uM. 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 10 units of restriction endonuclease for 4 hours. |
| Blue/White Cloning Assay | The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected. |
| Enzyme Activity | |
| Methylation Effect | Bpu1102I cuts GCTNAGm5C. |
| Stability during Prolonged Incubation | A minimum of 0.1unit 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. |
| |
| Compatible Ends: |
| Bpu10I, Eco81I, Hpy3I |
| Number of Recognition Sites in DNA | Lambda: 6 |
| PhiX174: 0 |
| M13mp18/19: 0 |
| pBR322: 0 |
| pUC18/19: 0 |
| pUC57: 0 |
| pTZ19R/U: 0 |
| pBluescriptIIKS(-/+): 0 |
| pBluescriptIISK(-/+): 0 |
| pACYC177: 0 |
| pACYC184: 0 |
| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
