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 si...

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Bibliographic Details
Published in:Russian journal of bioorganic chemistry 2021, Vol.47 (3), p.734-740
Main Authors: Brylev, V. A., Lysenko, I. L., Kokin, E. A., Martynenko-Makaev, Y. V., Ryazantsev, D. Y., Shmanai, V. V., Korshun, V. A.
Format: Article
Language:English
Subjects:
Biochemistry
Biomedical and Life Sciences
Biomedicine
Bioorganic Chemistry
Chemical reactions
Chemical synthesis
Dyes
Fluorescein
Life Sciences
Organic Chemistry
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Summary:— 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.
ISSN:1068-1620
1608-330X
DOI:10.1134/S1068162021030055