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A single-molecule primer extension assay. (A) Under an applied force of ∼3 pN, ssDNA is entropically collapsed, whereas dsDNA is extended to nearly its crystallographic length. Primer extension results in motion of tethered beads in the direction of flow. (B) Representative trajectories of synthesis by Pol III and Pol IV on individual DNA molecules (Fig. S1 E and F). (C and D) Processivity distributions for (C) Pol III (5 nM) and (D) Pol IV (30 nM); values represent means ± SEMs.
