 | Source: A Streptomyces sp. strain that carries the cloned Nae I gene from Nocardia aerocolonigenes.
Buffer supplied: 10x B1 (incl. 10x BSA).
Substrate for unit definition: pBR322 (4 sites).
Reaction conditions: 10 mM Tris-HCl (pH 7.9 @ 25°C), 10 mM MgCl2, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37°C. | | Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml BSA and 50 % glycerol. Store at -20°C.
Ligation and recutting: After ten-fold overdigestion with Nae I, >80 % of the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 65°C for 20 minutes.
Note: Nae I exhibits site preferences. Blocked by overlapping dcm methylation. | |
