| 用地高辛配基标记DNA探针及其应用 |
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| 引用本文： | 刘国仰,刘妙良.用地高辛配基标记DNA探针及其应用[J].中国医学科学院学报,1992,14(6):451-455. |
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| 作者姓名： | 刘国仰 刘妙良 |
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| 作者单位： | 中国医学科学院基础医学研究所 北京
(刘国仰，刘妙良，刘翀)，中国医学科学院基础医学研究所 北京(程在玉) |
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| 摘 要： | 核酸杂交技术中,同位素标记DNA探针是最常用的技术,其优越性是检测灵敏度高,但同时有很多缺点。80年代初,Langer等开始用生物素标记多核苷酸探针。随着方法的不断完善,近年来已用于基因诊断、定位及表达等研究。本文报道一种新的非同位素标记技术,即用地高辛配基(digoxigenin-dUTP)标记DNA探针,其原理是具有类固醇半抗原性质的地高辛配基,通过随机引物(random pr-imer法)掺入到DNA上,与靶DNA进行碱基互补同源序列结合形成杂交产物,然后通过酶联免疫法与
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| 关 键 词： | 地高辛 DNA探针 标记 基因诊断 |
DNA probe labeling with digoxigenin-dUTP and its application in gene diagnosis] |
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| G Liu.DNA probe labeling with digoxigenin-dUTP and its application in gene diagnosis][J].Acta Academiae Medicinae Sinicae,1992,14(6):451-455. |
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| Authors: | G Liu |
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| Institution: | Institute of Basic Medical Sciences, Beijing. |
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| Abstract: | In this paper, DNA probe labeling by the randomly primed incorporation of digoxigenin-dUTP is reported. The sensitivity of color reaction and hybridization were 32 fg and 200 fg, respectively, and both were specific for the target. Single-copy and multi-copy gene fragments among 2 micrograms human genomic DNA were detected by beta IVS II, Fr 3-42 and 3'HVR labeled with digoxigenin-dUTP. The results were consistent with a radioactive control assay. This method has been successfully used in the gene diagnosis of adult polycystic kidney disease. |
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