 | Source: Thermus aquaticus YT I.
Buffer supplied: 10x Taq I (incl. 10x BSA).
Substrate for unit definition: λ DNA, dam- (121 sites).
Reaction conditions: 100 mM KCl, 20 mM Tris-HCl (pH 8.5 @ 25°C), 3 mM MgCl2, 0.004% Triton X-100, 100 µg/ml BSA. Incubate at 65°C. | | Storage buffer: 300 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 500 µg/ml BSA and 50 % glycerol. Store at -20°C.
Ligation and recutting: After ten-fold overdigestion with Taq I, >90 % of the DNA fragments can be ligated and recut with this enzyme.
Heat inactivation: 80°C for 20 minutes.
Note: Taq I is blocked by overlapping dam methylation. Incubation without BSA results in 50 % activity. | |
