 | Source: Neisseria mucosa heidelbergensis (ATCC 25999).
Buffer supplied: 10x B5 (incl. 10x BSA).
Substrate for unit definition: λ DNA (Hind III digest).
Reaction conditions: 50 mM potassium acetate, 20 mM Tris-acetate (pH 7.9 @ 25°C), 10 mM magnesium acetate, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37°C.
Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml BSA and 50 % glycerol. Store at -20°C. | | Ligation and recutting: After 100-fold overdigestion with Nhe I, > 98 % of the DNA fragments can be ligated and recut.
Note: Activity inhibited by salt concentrations > 100mM. Cleaves to leave a 5' CTAG extension which can be efficiently ligated to DNA fragments generated by Avr II, Spe I, or Xba I.
Star activity: Large excess of the enzyme may results in the appearance of star activity.
Heat inactivation: 65°C for 20 minutes. | |
