| 5'-C C T N N^N N N A G G-3' |
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| 3'-G G A N N N^N N T C C-5' |
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| Catalog # | X0960 |
| Source | Xanthobacter agilis Vs18-132 |
| Supplied With | R1625: Restriction Enzyme Buffer A, 10X; (1X compostition after dilution)-33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. |
| R1625-04 | Restriction Enzyme Buffer E, 10X: 10mM Tris-HCl, pH 8.5,10mM magnesium chloride, 100mM potassium chloride, 0.1mg/ml BSA |
| 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 at 25°C), 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. |
| Storage Buffer | 10mM Tris-HCl (pH 7.5 at 25°C), 100mM potassium chloride, 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 XagI. |
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| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with XagI, approximately 80% of the DNA fragments can be ligated in the reaction mixture containing 20–40u of T4 DNA Ligase/1ug of fragments and 10% of PEG at a 5'-termini concentration of 0.1µM. More than 95% of these can be recut. |
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| 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. |
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| 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. |
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| Stability during Prolonged Incubation | A minimum of 0.1 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
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| Thermal Inactivation | Enzyme is inactivated by incubation at 65°C for 20min. |
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| Digestion of Agarose-embedded DNA | A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
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| Number of Recognition Sites in DNA | |
| Lambda | 9 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 1 |
| pUC18/19 | 0 |
| pUC57 | 0 |
| pTZ19R/U | 0 |
| pBluescriptIIKS(-/+) | 0 |
| pBluescriptIISK(-/+) | 0 |
| pACYC177 | 1 |
| pACYC184 | 1 |
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| 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. |
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