| 5'-G G G C C^C-3' |
|
| 3'-C^C C G G G-5' |
|
| Catalog # | A2298-63 |
| Concentration | 10u/ul |
| Source | Acetobacter pasteurianus |
| Buffer | 10mM Tris-HCl (pH 7.5), 10mM MgCl2 and 0.1mg/ml BSA. Incubate at 37°C. (Incubation at 30°C results in a 2-fold increase in activity.) |
| 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 7.4 at 25°C), 50mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. |
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with ApaI. |
| Ligation/Recutting Assay After 50-fold overdigestion (3u/ug DNA x 17 hours) with ApaI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.02uM. 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 0.3% of white colonies were detected. |
| Methylation Effects | Sequence GGGCCm5C is cleaved approximately 150 times slower than the unmethylated one. Overlapping Dcm or CG methylation may influence DNA cleavage. |
| Stability during Prolonged Incubation | A minimum of 0.2units of enzyme is required for complete digestion of 1ug of DNA in 16 hours at 37°C. |
| Thermal Inactivation | Enzyme is inactivated by incubation at 65°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 | GGGCC^C–BseSI, GGGCC^C–Eco24I, GGGCC^C–SduI |
| 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. |
| 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. |
