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二核苷酸重复多态性的非同位素检测及其在基因诊断中的应用

Non-radioactive Detection of Dinucleotide Repeat Polymorphisms and Its Application to Genetic Diagnosis

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【作者】 黄晓明胡向阳宗卉高翼之

【Author】 Huang Xiaoming; Hu Xiangyang; Zong Hui; Gao Yizhi(Division of Molecular Biology,Institute of Medical Science. Nanjing Railway Medical College, NanJing 210009)

【机构】 南京铁道医学院科研所医学分子生物学室

【摘要】 本文报道了一种检测二核苷酸重复多态性的简便的非同位素法,利用重复序列两侧的特异引物进行PCR扩增,产生的等位片段在薄层变性聚丙烯酰胺凝胶电泳上分离,再用灵敏的银染法显色。该方法不需要标记PCR产物,简便、快速,分辨率可达1bp,并可用多对引物同时进行多重PCR分析。用此方法对DMD家系成员dystrophin基因的5'-脑型外显子上游区和3'-非翻译区的两个(CA)n位点进行了扩增片段长度多态性分析,表明它是连锁分析和基因诊断的一种方便有效的新技术。

【Abstract】 We have developed a rapid and non-radioactive procedure for detecting dinucleotide repeat polymorphisms.Specific primers flanking the repeats were used in thepolymerase chain reaction(PCR).The generated allelic fragments were resolved bythin-layer denaturing polyacrylamide gel electrophoresis and visualized by improvedsilver staining. This method is simple, fast and without labelling of PCR products.Resolution of 1 bp can be easily obtained. It can also be used in multiplex PCR analysis of(CA)n loci. This method will be a valuable tool for linkage analysis andDNA diagnosis as demonstrated by its successful application to the CA dinucieotiderepeat haplotying of a DMD family.Received October 29.1993

  • 【文献出处】 遗传学报 ,ACTA GENETICA SINICA , 编辑部邮箱 ,1995年02期
  • 【分类号】R394
  • 【被引频次】29
  • 【下载频次】49
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