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A Two-Step Method for smFRET Data Analysis

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posted on 05.07.2016, 00:00 by Jixin Chen, Joseph R. Pyle, Kurt Waldo Sy Piecco, Anatoly B. Kolomeisky, Christy F. Landes
We demonstrate a two-step data analysis method to increase the accuracy of single-molecule Förster Resonance Energy Transfer (smFRET) experiments. Most current smFRET studies are at a time resolution on the millisecond level. When the system also contains molecular dynamics on the millisecond level, simulations show that large errors are present (e.g., > 40%) because false state assignment becomes significant during data analysis. We introduce and confirm an additional step after normal smFRET data analysis that is able to reduce the error (e.g., < 10%). The idea is to use Monte Carlo simulation to search ideal smFRET trajectories and compare them to the experimental data. Using a mathematical model, we are able to find the matches between these two sets, and back guess the hidden rate constants in the experimental results.