Figure 5. Ligation kinetics.

A) Ligation reactions were performed at 20°C using 3.3 µM ³²P-labeled dXHr8, 4.1 µM capture oligo and 4.1 µM of the Watson-Crick-matched DNA bridging template strands. Squares, ligated product; triangles, adenylated intermediate. Identities of the tethered nucleotide (asterisks) and bridges (in parentheses) are indicated. Values of kinetic rate constants k₂ (5′ DNA adenylation) and k₃ (gap-sealing) are given in the bottom right panel. B) Ligation reactions for tethered “universal” nucleoside, 5-nitroindole (dNI), were performed under the same conditions as in (A) using ³²P-labeled dNIHr8 in the presence of the indicated bridging template. Filled symbols, adenylate; open symbols, ligated product.