 | Source: Escherichia coli, J62plg 74.
Buffer supplied: 10x B2 (incl. 10x BSA).
Substrate for unit definition: λ DNA (21 sites).
Reaction conditions: 50 mM NaCl, 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 20-fold overdigestion with EcoR V, >95 % 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: 80°C for 20 minutes. | |
