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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'
Xanthobacter agilis Vs18-132
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
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.
10mM Tris-HCl (pH 7.4 at 25°C), 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
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:
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.