Table 4. Results of n and n – 1 mismatch investigation under optimized conditions.

The data show percent product formation and percent intermediate formation (in parentheses). The horizontal heading denotes template used and the vertical heading denotes the new ligated product. See Figure 1 for naming conventions of acceptors, donors and templates. Dark gray background denotes completely matched cases. All reactions were run at 22°C for 18 h. Reaction conditions were as follows: 30 mM Tris–HCl (pH 7.5), 10 mM DTT, 10 mM MgCl₂, 10 µM ATP, 20% DMSO, 2.5 pmol acceptor, 70 fmol donor and 3.125 pmol template.