Molecular Beacon DNA Probes with Fluorescein Bifluorophore

An azido-derivative of a fluorescein bifluorophore was obtained and used for the synthesis of “molecular beacon”-type oligonucleotide fluorogenic probes for RT-PCR. Eight probe variants were synthesized based on an optimized sequence: with one or two quencher residues at the 3'-end, with a sing...

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Autores principales: Brylev, V. A., Lysenko, I. L., Kokin, E. A., Martynenko-Makaev, Y. V., Ryazantsev, D. Y., Shmanai, V. V., Korshun, V. A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Pleiades Publishing 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8193957/
https://www.ncbi.nlm.nih.gov/pubmed/34149274
http://dx.doi.org/10.1134/S1068162021030055
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author Brylev, V. A.
Lysenko, I. L.
Kokin, E. A.
Martynenko-Makaev, Y. V.
Ryazantsev, D. Y.
Shmanai, V. V.
Korshun, V. A.
author_facet Brylev, V. A.
Lysenko, I. L.
Kokin, E. A.
Martynenko-Makaev, Y. V.
Ryazantsev, D. Y.
Shmanai, V. V.
Korshun, V. A.
author_sort Brylev, V. A.
collection PubMed
description An azido-derivative of a fluorescein bifluorophore was obtained and used for the synthesis of “molecular beacon”-type oligonucleotide fluorogenic probes for RT-PCR. Eight probe variants were synthesized based on an optimized sequence: with one or two quencher residues at the 3'-end, with a single or bifluorophore fluorescein label attached to 5'-end using modifying phosphoramidites (short linker) or “click reaction” (long linker). Comparison of probes in RT-PCR showed that probes with a doubled quencher (single fluorescein on a short linker) and doubled dye on a short linker (single dye) are somewhat superior in sensitivity to a standard probe (single quencher, single dye on a short linker) by the value of ΔC(t) = 1–2. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1134/S1068162021030055.
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spelling pubmed-81939572021-06-15 Molecular Beacon DNA Probes with Fluorescein Bifluorophore Brylev, V. A. Lysenko, I. L. Kokin, E. A. Martynenko-Makaev, Y. V. Ryazantsev, D. Y. Shmanai, V. V. Korshun, V. A. Russ J Bioorg Chem Article An azido-derivative of a fluorescein bifluorophore was obtained and used for the synthesis of “molecular beacon”-type oligonucleotide fluorogenic probes for RT-PCR. Eight probe variants were synthesized based on an optimized sequence: with one or two quencher residues at the 3'-end, with a single or bifluorophore fluorescein label attached to 5'-end using modifying phosphoramidites (short linker) or “click reaction” (long linker). Comparison of probes in RT-PCR showed that probes with a doubled quencher (single fluorescein on a short linker) and doubled dye on a short linker (single dye) are somewhat superior in sensitivity to a standard probe (single quencher, single dye on a short linker) by the value of ΔC(t) = 1–2. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1134/S1068162021030055. Pleiades Publishing 2021-06-11 2021 /pmc/articles/PMC8193957/ /pubmed/34149274 http://dx.doi.org/10.1134/S1068162021030055 Text en © The Author(s) 2021, ISSN 1068-1620, Russian Journal of Bioorganic Chemistry, 2021, Vol. 47, No. 3, pp. 734–740. © The Author(s), 2021. This article is an open access publication.Russian Text © The Author(s), 2021, published in Bioorganicheskaya Khimiya, 2021, Vol. 47, No. 3, pp. 365–372. https://creativecommons.org/licenses/by/4.0/Open Access.This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Brylev, V. A.
Lysenko, I. L.
Kokin, E. A.
Martynenko-Makaev, Y. V.
Ryazantsev, D. Y.
Shmanai, V. V.
Korshun, V. A.
Molecular Beacon DNA Probes with Fluorescein Bifluorophore
title Molecular Beacon DNA Probes with Fluorescein Bifluorophore
title_full Molecular Beacon DNA Probes with Fluorescein Bifluorophore
title_fullStr Molecular Beacon DNA Probes with Fluorescein Bifluorophore
title_full_unstemmed Molecular Beacon DNA Probes with Fluorescein Bifluorophore
title_short Molecular Beacon DNA Probes with Fluorescein Bifluorophore
title_sort molecular beacon dna probes with fluorescein bifluorophore
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8193957/
https://www.ncbi.nlm.nih.gov/pubmed/34149274
http://dx.doi.org/10.1134/S1068162021030055
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