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Method of tritium labeling oligonucleotide

机译:oligonucleotide标记寡核苷酸的方法

摘要

The present invention comprises a novel method of incorporating a tritium label at one or more predetermined sites within an oligonucleotide. In particular, the method comprises contacting a nascent, support-bound oligonucleotide having a free 5' hydroxyl group with a suitable oxidizing agent to oxidize the alcohol to an aldehyde, followed by reducing the aldehyde thereby formed with a suitable tritium labeled reducing agent such as [.sup.3 H]NaBH.sub.4 to yield the 5' terminal alcohol with a 5' tritium label. Normal automated synthesis can then be continued to yield the oligonucleotide of desired length having the tritium label in the desired location. The oligonucleotides thereby produced have higher specific activity than those previously known in the art. According, in a second aspect, the present invention provides oligonucleotides having high specific acitivity. The oligonucleotides of the present invention are useful for determining the pharmacokinetics and biodistribution of their non-radiolabeled counterparts, both in vitro and in vivo.
机译:本发明包括一种在method寡核苷酸内一个或多个预定位点掺入a标记的新颖方法。特别地,该方法包括使具有游离5'羟基的新生的,与载体结合的寡核苷酸与合适的氧化剂接触,以将醇氧化为醛,然后还原醛,从而与合适的tri标记的还原剂如[3 H] NaBH.sub.4生成带有5'label标记的5'末端醇。然后可以继续正常的自动合成以产生具有所需长度的寡核苷酸,该寡核苷酸在所需位置具有having标记。由此产生的寡核苷酸比本领域先前已知的具有更高的比活性。根据第二方面,本发明提供了具有高比活性的寡核苷酸。本发明的寡核苷酸可用于在体外和体内测定其非放射性标记的对应物的药代动力学和生物分布。

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