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PYRENE-MODIFIED RNA, AND ASSAY OF RNA

机译:Y修饰的RNA和RNA测定

摘要

PROBLEM TO BE SOLVED: To obtain a new RNA containing pyrene-incorporated base(s) in its constituent base sequence, and emitting intense fluorescence through its hybridization with the corresponding complementary strand, therefore assayable in its reaction in a homogenous system by the aid of measuring the above fluorescence. ;SOLUTION: This new RNA is such one as to contain pyrene-incorporated base(s) in its constituent base sequence, emit intense fluorescence when hybridized with the corresponding complementary strand, and therefore be assayable in a homogeneous system. The assay technique is useful in e.g. in-situ detection, variant detection. This pyrene-modified RNA is obtained according to the following procedure: a reaction is carried out between uridine with the hydroxyl groups on 3'- and 5'-sites protected with trityl groups and 1-chloromethylpyrene followed by deprotection to covert into 2'-O-(1-pyrenylmethyl)uridine, with which a 4,4'-dimethoxytritylpyrene-modified uridine-phosphorodiamidate reagent is prepared, and subsequently, using this reagent, a pyrene-modified uridine is incorporated into RNA by using an automatic DNA synthesizer.;COPYRIGHT: (C)2000,JPO
机译:要解决的问题:要获得一种新的RNA,该RNA的组成碱基序列中包含掺入base的碱基,并通过与相应互补链的杂交而发出强烈的荧光,因此可以借助以下方法在同质系统中的反应中进行测定测量上述荧光。 ;解决方案:这种新RNA的组成碱基序列中含有掺入pyr的碱基,与相应的互补链杂交后发出强烈的荧光,因此可以在均质系统中进行测定。该测定技术可用于例如原位检测,变异检测。该pyr修饰的RNA是根据以下步骤获得的:尿苷与在三苯甲基保护的3'-和5'-位上的羟基和1-氯甲基py之间进行反应,然后脱保护成2'-制备O-(1-吡啶基甲基)尿苷,用其制备4,4'-二甲氧基三苯并lp改性的尿苷-磷酸二氨基甲酸酯试剂,随后使用该试剂,通过使用自动DNA合成仪将a改性的尿苷掺入RNA。 ;版权:(C)2000,日本特许厅

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