Significance of methods for purification of oligodeoxyribonucleotide probes for the efficiency of gene diagnosis by real-time PCR |
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Authors: | O N Tatarinova T N Lukyanova M A Zaitseva K Yu Veremeev V A Karpov A N Chuvilin D D Petrunin G E Pozmogova |
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Institution: | (1) Institute of Physicochemical Medicine, Russian Ministry of Health, Moscow, Russia;(2) Litekh Company, Moscow, Russia |
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Abstract: | Analysis of the use of real-time PCR with fluorescent registration of results for gene diagnosis of infectious diseases showed
that the sensitivity and reliability of quantitative evaluation of DNA targets directly depended on the method of purification
of oligonucleotide probes. Chromatographic behavior of synthetic probes carrying various fluorophores and fluorescence quenchers
was analyzed. Approaches to optimization of purification methods are proposed enabling elimination of previously undetectable
admixtures. The importance of these studies is explained by the need in extending the armory of methods for the development
and production of diagnosticums for detection of infectious and hereditary diseases, identification of genetically modified
organisms, and for a wide spectrum of research in molecular biology and medicine.
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Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 3, pp. 280–284, March, 2008 |
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Keywords: | oligodeoxyribonucleotide probes polymerase chain reaction in the real-time mode gene diagnosis high performance liquid chromatography mass spectrometry |
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