Fig. 5. Measured anneal switch temperature as a function of ratio of l-DNA quencher strand to fluorophore strand (red circles, mean ± s.d., N = 4). Eqn (3) with ΔH and ΔS calculated from the experimental data shows the overall trend of the data (dashed blue line). The 2.5 ratio used for the total concentration experiments also results in a switch temperature of 59.6 °C (black dashed line and arrow). The limit of the switch temperature as ([C₁]/[C₂]) approaches infinity for a fixed total concentration of 500 nM is 60.3 °C (dashed orange line).