 | Source: Nocardia otitidis-caviarum.
Buffer supplied: 10x Not I Buffer (incl. 10x BSA).
Substrate for unit definition: Adenovirus-2 DNA (7 sites).
Reaction conditions: 100 mM NaCl, 50 mM Tris-HCl (pH 7.9 @ 25°C), 5 mM MgCl2, 1 mM DTT, 100 µg/ml BSA. Incubate at 37°C.
Storage buffer: 500 mM KCl, 10 mM Tris-HCl (pH 7.4), 1 mM DTT, 0.1 mM EDTA, 0.1% Triton X-100, 500 µg/ml BSA and 50% glycerol. Store at -20°C. | | Ligation and recutting: After 25-fold overdigestion with Not I, >98% of the DNA fragments can be ligated and recut with this enzyme.
Note: Supercoiled plasmids may require up to 5-fold more Not I for complete digestion than linear DNA.
Heat inactivation: 65°C for 20 minutes. | |
