FIGURE 6.

Quantification of species from AFM images of reactions containing lds_het–ins and css_hom DNA. (A) Scheme of RecA-mediated recombination reaction between css_hom (open circle) and lds_het–ins (internal black square and a gray square at 5′-end) substrates, showing the jm intermediates and the hypothetical final nc product (in gray). The 3276-nt css_hom DNA (10 μM in nt) was preincubated with SsbA, RecO and RecA (60 min, 37°C) in buffer A with 5 mM ATP. Then, the 3353-bp lds_het–ins DNA was added without (B) or with MutS (C). Quantification of the different DNA species observed in the absence of MutS (n = 293 molecules) (D) or in the presence of MutS (n = 419 molecules) (E). Quantification only includes dsDNA molecules with the original length and ssDNA molecules with a volume corresponding to the original length. Agg, css_hom with lds_het–ins DNA at one end and internal regions or more complex aggregates. Arrows point to examples of the different species (black: ssDNA; red: dsDNA; light blue: nicked circular; green: Agg).