Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification
Objectives. To study the substrate properties of Cy5-labeled deoxynucleoside triphosphates of various natures (dU and dC) in the process of incorporation in the DNA chain during recombinase polymerase amplification (RPA).Methods. The work used the real-time RPA method. The method of horizontal elect...
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| Format: | Article |
| Language: | Russian |
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MIREA - Russian Technological University
2024-11-01
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| Series: | Тонкие химические технологии |
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| Online Access: | https://www.finechem-mirea.ru/jour/article/view/2158 |
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| author | A. S. Epifanov V. E. Shershov S. A. Surzhikov A. V. Chudinov S. A. Lapa |
| author_facet | A. S. Epifanov V. E. Shershov S. A. Surzhikov A. V. Chudinov S. A. Lapa |
| author_sort | A. S. Epifanov |
| collection | DOAJ |
| description | Objectives. To study the substrate properties of Cy5-labeled deoxynucleoside triphosphates of various natures (dU and dC) in the process of incorporation in the DNA chain during recombinase polymerase amplification (RPA).Methods. The work used the real-time RPA method. The method of horizontal electrophoresis was used to control the quality of the amplification products obtained.Results. The influence of the fluorophore structure and linker lengths on the substrate properties for deoxynucleoside triphosphates Cy5-dUTP and Cy5-dCTP was studied. The following values of the substrate efficiency parameters were determined: amplification efficiency (kinetic indicator), normalized product yield, and embedding coefficient.Conclusions. Modified deoxynucleoside triphosphates (dNTP) with long linkers between the fluorophore and the nitrogenous base, as well as between the quaternary ammonium group and the second heterocycle of the fluorophore, showed greater substrate efficiency than fluorescently labeled dNTP with short linkers. The modified dU in each pair demonstrated greater substrate efficiency compared to the modified dC. |
| format | Article |
| id | doaj-art-dcdb906d2a9f4d908ffe312c79ec114e |
| institution | Kabale University |
| issn | 2410-6593 2686-7575 |
| language | Russian |
| publishDate | 2024-11-01 |
| publisher | MIREA - Russian Technological University |
| record_format | Article |
| series | Тонкие химические технологии |
| spelling | doaj-art-dcdb906d2a9f4d908ffe312c79ec114e2025-08-20T03:56:32ZrusMIREA - Russian Technological UniversityТонкие химические технологии2410-65932686-75752024-11-0119540841710.32362/2410-6593-2024-19-5-408-4171776Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplificationA. S. Epifanov0V. E. Shershov1S. A. Surzhikov2A. V. Chudinov3S. A. Lapa4Engelhardt Institute of Molecular Biology, Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesObjectives. To study the substrate properties of Cy5-labeled deoxynucleoside triphosphates of various natures (dU and dC) in the process of incorporation in the DNA chain during recombinase polymerase amplification (RPA).Methods. The work used the real-time RPA method. The method of horizontal electrophoresis was used to control the quality of the amplification products obtained.Results. The influence of the fluorophore structure and linker lengths on the substrate properties for deoxynucleoside triphosphates Cy5-dUTP and Cy5-dCTP was studied. The following values of the substrate efficiency parameters were determined: amplification efficiency (kinetic indicator), normalized product yield, and embedding coefficient.Conclusions. Modified deoxynucleoside triphosphates (dNTP) with long linkers between the fluorophore and the nitrogenous base, as well as between the quaternary ammonium group and the second heterocycle of the fluorophore, showed greater substrate efficiency than fluorescently labeled dNTP with short linkers. The modified dU in each pair demonstrated greater substrate efficiency compared to the modified dC.https://www.finechem-mirea.ru/jour/article/view/2158recombinase polymerase amplificationfluorescently labeled deoxynucleoside triphosphates |
| spellingShingle | A. S. Epifanov V. E. Shershov S. A. Surzhikov A. V. Chudinov S. A. Lapa Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification Тонкие химические технологии recombinase polymerase amplification fluorescently labeled deoxynucleoside triphosphates |
| title | Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification |
| title_full | Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification |
| title_fullStr | Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification |
| title_full_unstemmed | Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification |
| title_short | Investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification |
| title_sort | investigation of the substrate properties of fluorescently labeled pyrimidine triphosphates in recombinase polymerase amplification |
| topic | recombinase polymerase amplification fluorescently labeled deoxynucleoside triphosphates |
| url | https://www.finechem-mirea.ru/jour/article/view/2158 |
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