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3 '- amino - or thiolmodifizierte, fluoreszenzfarbstoff - coupled nukleoside, nucleotides and oligonucleotides and a method for the production and their use

机译:3'-氨基-或巯基修饰的菲尔泽夫,荧光素-法布斯托夫-偶联的核苷,核苷酸和寡核苷酸及其生产和使用方法

摘要

The OH group located in the 3' and/or 2' position of a nucleoside, nucleotide or oligonucleotide is derivatised to an amino or thiol group and subsequently a fluorescent dye is coupled thereto. The resulting 3'- and/or 2'-amino- and thiol-modified nucleosides, nucleotides and oligonucleotides can then be used for the synthesis of items in the presence of a template strand or of oligonucleotides, and for the detection of genetic material. They offer the advantage that the fluorescent label is no longer attached to the 5' end of the oligonucleotide or to the nucleobase and thus does not have to be introduced during the chemical synthesis as in labelling techniques hitherto disclosed, while the known and conventional methods have the disadvantage that only some polymerases can be employed for the synthesis, the acceptance of the triphosphates by the polymerases falls and, in addition, a large substrate excess is necessary.
机译:位于核苷,核苷酸或寡核苷酸的3'和/或2'位置的OH基团被衍生为氨基或硫醇基团,随后将荧光染料偶联至其上。然后可以将所得的3'-和/或2'-氨基和硫醇修饰的核苷,核苷酸和寡核苷酸用于在模板链或寡核苷酸存在下的合成,以及用于检测遗传物质。它们提供的优点是荧光标记不再附着于寡核苷酸的5'端或核碱基,因此不必像迄今为止公开的标记技术那样在化学合成过程中引入,缺点是只能使用某些聚合酶进行合成,聚合酶对三磷酸酯的接受度下降,此外,需要大量的底物过量。

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