NdeI

5'-C A^T A T G-3' \n3'-G T A T^A C-5'\n\nBuffers:\nR1625-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 37 degrees C.\n\nR1625-03-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 37 degrees C.\n\nOverdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with NdeI. \n\nLigation/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. \n\nLabeled 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.\n\nStability 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 37 degrees C. \n\nThermal Inactivation: Ndel is inactivated by incubation at 65 degrees C for 20min. \n\nDigestion 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. \n\nCompatible Ends: Csp6I, FspBI, Tru1I, VspI \n\nNumber of Recognition Sites in DNA:\nLambda: 7\nPhiX174: 0\nM13mp18/19: 3\npBR322: 1\npUC18/19: 1\npUC57: 1\npTZ19R/U: 0\n\nUnit Definition: \nOne unit is defined as the amount of Ndel required to digest 1ug of lambda DNA in 1 hour at 37 degrees C in 50ul of assay buffer.\n\nDilution Buffer:\n10mM Tris-HCl, pH 7.4 at RT, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.\n\nEnzyme Activity in Buffer A (R1625)/Buffer D (R1625-03):\nR1625-0-20%\nR1625 (2X)-50-100%\nR1625-03-100%

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