Computational methods for sequence mapping of large combinatorial libraries and deduced sequence signatures |
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Authors: | Abécassis Valérie Aggerbeck Lawrence Truan Gilles Pompon Denis |
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Institution: | Centre National de la Recherche Scientifique, UPR 2167, Gifsur-Yvette, France. |
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Abstract: | Here we describe a computational approach for the high-throughput sequence mapping of combinatorial libraries obtained by DNA shuffling. Original algorithms and their software implementation were developed for the automated and reliable analysis of hybridization data of differentially labeled oligonucleotide probes with PCR products spotted on DNA microarrays. This novel approach allows a context-dependent sequence attribution tolerant to fluctuations in experimental conditions and is well adapted to hybridization signals of variable qualities resulting from high-throughput PCR amplification from colonies. In addition, the analysis permits the calculation of sequence signatures that are characteristic of combinatorial library structure, defects, and diversity. The approach is of interest for the characterization and the equalization (library reduction to nonredundant structures) of combinatorial libraries involved in directed evolution and could be extrapolated to high-throughput polymorphism analysis. |
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