Methods are provided for post incorporation labeling of a nucleic acid, including for example cRNA, labeled with nucleotide analogs having a formula selected from the group consisting of
wherein A is H or a functional group that permits the attachment of the nucleic acid labeling compound to a nucleic acid; Y and Z are independently H or OH; L is linker group; and P is a connecting group. After incorporation of the nucleic acid (including cRNA) with the above nucleotide analogs, the nucleic acid is labeled with a detectable group reagent wherein said detetable group reagent comprises a chemical moiety which is capable of specifically reacting with said P group to allow coupling of the detectable group to said primed cRNA. Compounds comprising nucleotide analogs are also presented in accordance with the present invention. Methods are also presented for incorporating these compounds into nucleic acids and subsequently labeling the incorporated nucleotide analog with detectable moiety reagent.
本发明提供了用核苷酸类似物对核酸(包括例如 cRNA)进行掺入后标记的方法,核苷酸类似物的分子式选自由以下组成的组
其中 A 是 H 或允许核酸标记化合物附着到核酸上的官能团;Y 和 Z 独立地是 H 或 OH;L 是连接基团;P 是连接基团。在核酸(包括 cRNA)与上述核苷酸类似物结合后,用可检测基团试剂标记核酸,其中所述可检测基团试剂包括能与所述 P 基团发生特异性反应的
化学分子,以便将可检测基团与所述引物 cRNA 偶联。根据本发明还提出了包含核苷酸类似物的化合物。还介绍了将这些化合物掺入核酸并随后用可检测的分子试剂标记掺入的核苷酸类似物的方法。