Figure EV4. Tandem‐target dwelltime and single‐target binding rate.

1. Dwelltime distributions for tandem‐target construct for each target separation, fitted with a double‐exponential decay function. Equation of the fit: y = A₁e^−t/τ1 + A₂e^−t/τ2. A values for each tandem‐target construct: d6 ‐ A₁ = 640.3, A₂ = 8.0; d9 ‐ A₁ = 654.7, A₂ = 8.8; d12 ‐ A₁ = 144.6, A₂ = 6.4; d23 – A₁ = 635.3 A₂ = 5.6. Average dwelltime for each construct: d6 ‐ τ_av = 12.3 s_; d9 ‐ τ_av = 6.9 s; d12 ‐ τ_av = 10.7 s; d23 ‐ τ_av = 7.6 s. Average dwelltimes were calculated using equation ${\mathrm{\tau }}_{\mathit{av}}=\frac{A_1{\mathrm{\tau }}_1^2+A_2{\mathrm{\tau }}_2^2}{A_1{\mathrm{\tau }}_1+A_2{\mathrm{\tau }}_2}$
2. Scatter plot showing the total binding rate (k_on) for each single‐target, multiple PAM construct. Binding rate stays constant with increasing number of adjacent PAMs. The values are averages of four measurements over 3 days. Error bars represent standard error of the mean. k_on was calculated using the equation k_on = 1/(CxΔτ_on) where C is the molarity of DNA target in the channel.