11 July 2001 Interpretation of the fluorescence image acquired for DNA sequencing
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Proceedings Volume 4515, Light and Optics in Biomedicine; (2001) https://doi.org/10.1117/12.432991
Event: Light and Optics in Biomedicine, 2000, Warsaw, Poland
Abstract
This paper is devoted to problems with reliability of the fluorescence analysis in investigating the DNA chains. Relationships between thermodynamic and optical properties of a given sequence are taken to consider modeling of a virtual light-directed library of oligonucleotides. Specificity of changes in optical density caused by specific interactions between pairs of DNA bases depends on the known nearest neighbor model which has been assumed here as the background. If to assume an integrated array of the known oligonucleotides (L-mers) which all have their own stable locations, after hybridization we may obtain a fluorescence result creating a matrix of micro-images; each one corresponds to the optical answer of a given oligonucleotide included in the investigated sequence spectrum. A presented concept of interpretation of fluorescence effects depends on relation between a given level of optical density and order of the bases included in oligonucleotides involved in hybridization procedure. The factors influencing fluorescence image attributes, including signal levels and stability as well as resolution and accuracy of sensing, are discussed. The obtained results of comparative computations made on a normal and simulated abnormal composition of a given human sequence are illustrated at the case that L = 6. Basing on consequences of changes in melting temperature, thermodynamic stability and levels of fluorescence light intensity, the optical spectra are presented as images made by sites of full hybridization put on a reference thermal map.
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Anna Cysewska-Sobusiak, Michal Boltrukiewicz, "Interpretation of the fluorescence image acquired for DNA sequencing", Proc. SPIE 4515, Light and Optics in Biomedicine, (11 July 2001); doi: 10.1117/12.432991; https://doi.org/10.1117/12.432991
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