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Control of DNA Strand Displacement Kinetics Using Toehold Exchange

https://doi.org/10.1021/ja906987s
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no DOI — not checkedOn the basis of the dependence ofkf1on the sequence composition of the toehold, we believe thatkf2is likely closest in value tokf1= 3 × 106M−1s−1for inputs X(m, n) with toeholds containing both strong G-C pairs and weak A-T pairs. However, we have no way of knowing the actual value ofkf2without performing a new series of experiments on theβmtoehold, similar to those shown in Figure3.
no DOI — not checkedTo take a simple example, consider when the invading and incumbent toeholds are both 4 nt but the branch migration domain is 1000 nt long. Molecules will spend a significant amount of time in a three-stranded complex, with the branch migration junction vacillating near the middle of the branch migration domain. Our model, with the 2/(b − m) correction, will account for the high multiplicity of branch migration microstates.
no DOI — not checkedODEs used for fittingk{m, n}simulated the two reactions X(m, n) + S → Y + L(m, n) and Y + R → F. The reverse reaction Y + L(m, n) → S + X(m, n) is not included because (1) it is desireable to fit one parameter at a time and (2) because the value of the reverse rate constantκ{n, m}is constrained poorly by data from experiments of the type shown in Figure3A. Furthermore, the concentration of the reporter R is in 3× excess over that of the substrate S, and the rate constant of the reaction between Y and R is high; consequently, it is expected that the reaction between Y and R will dominate that between Y and L(m, n). Fittingk{m, n}to a full model including the reverse reaction with its rate constantκ{m, n}set to the value predicted by our model yielded values very similar to those presented in the paper (difference of no more than 10%).
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