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5'-C A^T A T G-3'
3'-G T A T^A C-5'
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 37°C.
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 37°C.
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 37°C.
Thermal Inactivation: Ndel is inactivated by incubation at 65°C 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:
One unit is defined as the amount of Ndel 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 RT, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Enzyme Activity in Buffer A (R1625)/Buffer D (R1625-03):
Source: Neisseria denitrificans
Form: Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.