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FRET experiments in which donor (YO-PRO-1) and acceptor (TO-PRO-3) were cointercalated at varying ratios in a DNA 3WJ template. Legend lists YO-PRO-1 and TO-PRO-3 concentrations in micromolar (e.g. 14:1 = 14 μM YO-PRO-1 and 1 μM TO-PRO-3. (A) Experimental data acquired with excitation at 440 nm. [DNA 3WJ] = 1.0 μM. (B) Simulated data corresponding to FRET efficiencies determined based on quenching of donor from spectra in (A), as described in Materials and Methods section.
