Description: 5-T T T A A A-3 3-A A A T T T-5 Concentration: 10u/ul Source: Deinococcus radiophilus Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Form: Supplied as a liquid in 10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 0.15% Triton X-100 and 50% glycerol. Incubate at 37C Dilution Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. R1625 (Included Buffer): 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (for 100% DraI digestion). Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (20u/ug lambda DNA x 16 hours) with DraI. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with DraI, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.13uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide was observed after incubation with 10 units of DraI for 4 hours. Methylation Effect: DraI does not cut Dam, Dcm, CpG, EcoBl. Blocked by overlapping EcoKI methylation. Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min. 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 37C. Digestion of Agarose-embedded DNA: A minimum of 5 units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda: 13 PhiX174: 2 M13mp18/19: 5 pBR322: 3 pUC18/19: 3 pUC57: 3 Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Form:
Supplied as a liquid in 10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 0.15% Triton X-100 and 50% glycerol. Incubate at 37C
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