Brylev V A, Lysenko I L, Kokin E A, Martynenko-Makaev Y V, Ryazantsev D Y, Shmanai V V, Korshun V A
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia.
Institute of Physical Organic Chemistry of NAS Belarus, 220072 Minsk, Belarus.
Russ J Bioorg Chem. 2021;47(3):734-740. doi: 10.1134/S1068162021030055. Epub 2021 Jun 11.
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.
The online version contains supplementary material available at 10.1134/S1068162021030055.
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。
在线版本包含可在10.1134/S1068162021030055获取的补充材料。
