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Technical Data |
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X0960 |
XagI (EcoNI) |
1000u 5000u |
| Molecular Biology | Storage: -20°CShipping: Blue Ice |
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5'-C C T N N^N N N A G G-3' 3'-G G A N N N^N N T C C-5' 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. 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. 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. 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. Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20min. 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. 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 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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