Fourier analysis to measure diffusion coefficients and resolve mixtures on a continuous electrophoresis chip.

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Estevez-Torres A, Gosse C, Le Saux T, Allemand JF, Croquette V, Berthoumieux H, Lemarchand A, Jullien L

Fourier analysis to measure diffusion coefficients and resolve mixtures on a continuous electrophoresis chip.

Anal Chem. 2007 Nov 1;79(21):8222-31. Epub 2007 Sep 25.

PubMed ID

17892271 [ View in PubMed

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Abstract

We report a method to measure diffusion coefficients of fluorescent solutes in the 10(2)-10(6) Da molecular mass range in a glass-PDMS chip. Upon applying a permanent electric field, the solute is introduced through a narrow channel into a wide analysis chamber where it migrates along the injection axis and diffuses in two dimensions. The diffusion coefficient is extracted after 1D Fourier transform of the resulting stationary concentration pattern. Analysis is straightforward, requiring no numerical integration or velocity field simulation. The diffusion coefficients measured for fluorescein, rhodamine green-labeled oligonucleotides, and YOYO-1-stained dsDNA fragments agree with the literature values and with our own fluorescence correlation spectroscopy measurements. As shown for 151 and 1257 base pair dsDNA mixtures, the present method allows us to rely on diffusion to quantitatively characterize the nature and the composition of binary mixtures. In particular, we implement a DNA hybridization assay to illustrate the efficiency of the proposed protocol for library screening.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Fluorescein	DNA	Nucleotide	Humans	No	Other	Details