5-C A T A T G-3 3-G T A T A C-5 Source: Neisseria denitrificans Concentration: ~10u/ul Buffers: R1625-Restriction Enzyme Buffer A (double digests): 1X Buffer composition-Supplied as a liquid containing 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37C. R1625-03-Restriction Enzyme Buffer D (for 100% digestion): 1X Buffer composition-Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA. Incubate at 37C. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with NdeI. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with NdeI, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.1uM. 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. Stability during Prolonged Incubation: A minimum of 0.3 units of Ndel is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Thermal Inactivation: Ndel is inactivated by incubation at 65C for 20min. Digestion of Agarose-embedded DNA: A minimum of 5 units of Ndel is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Compatible Ends: Csp6I, FspBI, Tru1I, VspI Number of Recognition Sites in DNA: Lambda: 7 PhiX174: 0 M13mp18/19: 3 pBR322: 1 pUC18/19: 1 pUC57: 1 pTZ19R/U: 0 Unit Definition: One unit is defined as the amount of Ndel required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Applications: Molecular cloning Restriction site mapping Genotyping Southern blotting Restriction fragment length polymorphism (RFLP) SNP Dilution Buffer: 10mM Tris-HCl, pH 7.4 at RT, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Enzyme Activity in Buffer A (R1625)/Buffer D (R1625-03): R1625-0-20% R1625 (2X)-50-100% R1625-03-100% 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 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
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