Distribution analysis for single molecule FRET measurement.
A new numerical analysis method for experimental single-pair fluorescence resonance energy transfer (sp-FRET) data is proposed. In this method, every single data point was plotted in a style of a cumulative distribution function and dedicated to curve-fitting analysis, so that the analysis does not depend on bin size. A series of numerical simulations showed that this analysis has a more efficient and accurate resolvability of components than a fitting method based on Gaussian functions to a histogram plot. A simulated data based on experimental FRET distributions were also used to discuss the fitting errors of this method. The proposed method was applied to sp-FRET experiments of doubly dye-labeled double-strand DNA with a short sequence. Mixtures of up to three species were analyzed, and the contributions up to four subpopulations were successfully resolved.
Okamoto
Kenji
K
Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto, Kyoto 606-8502, Japan.
Terazima
Masahide
M
D000465:Algorithms; D001710:Biotin / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D003198:Computer Simulation; D004247:DNA / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000706:statistics & numerical data*; D005456:Fluorescent Dyes / Q000737:chemistry; D018613:Microscopy, Confocal; D011089:Polydeoxyribonucleotides / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry; D016008:Statistical Distributions
D002232:Carbocyanines; D005456:Fluorescent Dyes; D011089:Polydeoxyribonucleotides; D012235:Rhodamines; C085321:cyanine dye 5; C005358:tetramethylrhodamine; D001710:Biotin; D004247:DNA