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Method for labeling nucleotides by reacting a reactive platinum compound having two reactive moieties, a labeled moiety and another moiety of a spacer with a nucleotide

机译:通过使具有两个反应性部分,标记的部分和间隔基的另一个部分的反应性铂化合物与核苷酸反应来标记核苷酸的方法

摘要

A method for improved labeling of nucleotides comprised by the steps of: reacting a reactive moiety of a linker of the Formula (I), wherein X represents any stabilising bridge, A and B represents the same or different reacting moieties, with an electron donating moiety (aromatic amine or thiolate anion) of a spacer, which spacer comprises a chain having at least four atoms and at least one heteroatom (oxygen) in the chain, which spacer further comprises said electron donating moiety at one end of the chain and a reactive moiety at the other end of the chain; reacting the reactive moiety of said spacer with a label (biotin, avidin, streptavidin, digoxygenin or a functional equivalent thereof); .reacting the other reactive moiety of said linker with a nucleotide (adenine, thymidine, cytosine, guanine or uridine) A major advantage of this method is that all nucleotides can be labeled by the method described, whereas until now the attachment of a label was mostly restricted to one or certain nucleotides.
机译:一种改善核苷酸标记的方法,该方法包括以下步骤:使式(I)的连接体的反应性部分与供电子部分反应,其中X代表任何稳定桥,A和B代表相同或不同的反应部分。间隔基的(芳族胺或硫醇根阴离子),该间隔基包含在链中具有至少四个原子和至少一个杂原子(氧)的链,该间隔基还包含在链的一端的所述给电子部分和反应性链另一端的部分;使所述间隔物的反应性部分与标记物(生物素,亲和素,链霉亲和素,地高辛配基或其功能等同物)反应;用核苷酸(腺嘌呤,胸腺嘧啶,胞嘧啶,鸟嘌呤或尿苷)反应所述接头的另一个反应性部分,该方法的主要优点是所有核苷酸均可通过所述方法进行标记,而到目前为止,标记的附着是主要限于一个或某些核苷酸。

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