 | Source: Escherichia coli RY 13.
Buffer supplied: 10x EcoR I (incl. 10x BSA).
Substrate for unit definition: λ DNA (5 sites).
Reaction conditions: 50 mM NaCl, 100 mM Tris-HCl (pH 7.4 @ 25°C), 5 mM MgCl2, 0.025 % Triton X-100, 100 µg/ml BSA. Incubate at 37°C.
Storage buffer: 300 mM NaCl, 5 mM KPO4 (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 0.15 % Triton X-100, 200 µg/ml BSA and 50 % glycerol. Store at -20°C. | | Ligation and recutting: After 50-fold overdigestion with EcoR I, >98 % of the DNA fragments can be ligated and recut with this enzyme.
Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration >5 %, or pH >8.0 may result in star activity.
Heat inactivation: 65°C for 20 minutes. | |
