Significance of methods for purification of oligodeoxyribonucleotide probes for the efficiency of gene diagnosis by real-time PCR

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 beh...

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Bibliographic Details
Published in:Bulletin of experimental biology and medicine 2008-03, Vol.145 (3), p.312-316
Main Authors: Tatarinova, O. N., Lukyanova, T. N., Zaitseva, M. A., Veremeev, K. Yu, Karpov, V. A., Chuvilin, A. N., Petrunin, D. D., Pozmogova, G. E.
Format: Article
Language:English
Subjects:
Biomedical and Life Sciences
Biomedicine
Biophysics and Biochemistry
Cell Biology
Chromatography, High Pressure Liquid - methods
DNA - analysis
Humans
Internal Medicine
Laboratory Medicine
Oligonucleotide Probes - isolation & purification
Pathology
Polymerase Chain Reaction - methods
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Summary: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.
ISSN:0007-4888
1573-8221
DOI:10.1007/s10517-008-0078-6