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Journal Articles Electrophoresis Year : 2008

Quantitative predictions for DNA two-dimensional display according to size and nucleotide sequence composition

Abstract

2-D DNA display is a simple separation method that provides a fast and economical way of visualizing polymorphism and comparing genomes. The DNA fragments are separated first according to their size by standard gel electrophoresis and then according to their sequence composition using denaturing gradient gel electrophoresis. First developed by Fischer and Lerman (Cell 1979, 16, 191-200), this method has recently been used to distinguish strains within a bacterial species. The genomic restriction fragments are displayed as spots on a 2-D surface. Although most of the relevant physical mechanisms are understood, this technique is mostly empirical and remains essentially qualitative. In view of optimizing this procedure, we combine our understanding of the different physical mechanisms at play to develop a complete numerical model to predict the relative coordinates of the spots as a function of the corresponding DNA sequence and of the experimental conditions. We experimentally validate our model by predicting the outcome of a 2-D display of the phage genome. It thus becomes possible to optimize in silico the experimental parameters, to predict whether specific mutations as well as yet undescribed genetic polymorphisms can be resolved, and to assist in interpreting the experimental data.

Dates and versions

hal-00368941 , version 1 (18-03-2009)

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Jean-François Mercier, Christine Kingsburry, Gary W. Slater, Bénédicte Lafay. Quantitative predictions for DNA two-dimensional display according to size and nucleotide sequence composition. Electrophoresis, 2008, 29 (6), pp.1264 - 1272. ⟨10.1002/elps.200700407⟩. ⟨hal-00368941⟩
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