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含荧光素双荧光团的分子信标DNA探针。

Molecular Beacon DNA Probes with Fluorescein Bifluorophore.

作者信息

Brylev V A, Lysenko I L, Kokin E A, Martynenko-Makaev Y V, Ryazantsev D Y, Shmanai V V, Korshun V A

机构信息

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia.

Institute of Physical Organic Chemistry of NAS Belarus, 220072 Minsk, Belarus.

出版信息

Russ J Bioorg Chem. 2021;47(3):734-740. doi: 10.1134/S1068162021030055. Epub 2021 Jun 11.

Abstract

UNLABELLED

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 = 1-2.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1134/S1068162021030055.

摘要

未标注

获得了一种荧光素双荧光团的叠氮衍生物,并将其用于合成用于逆转录聚合酶链反应(RT-PCR)的“分子信标”型寡核苷酸荧光探针。基于优化序列合成了八个探针变体:在3'端带有一个或两个淬灭残基,使用修饰亚磷酰胺(短连接子)或“点击反应”(长连接子)在5'端连接单个或双荧光团荧光素标记。RT-PCR中探针的比较表明,具有双淬灭剂(短连接子上单个荧光素)和短连接子上双染料(单个染料)的探针在灵敏度上比标准探针(单个淬灭剂,短连接子上单个染料)略高,ΔC值为1-2。

补充信息

在线版本包含可在10.1134/S1068162021030055获取的补充材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e198/8193957/4176e7b12edd/11171_2021_8334_Fig1_HTML.jpg

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