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通过聚合酶链反应技术快速检测高变区。

Rapid detection of hypervariable regions by the polymerase chain reaction technique.

作者信息

Decorte R, Cuppens H, Marynen P, Cassiman J J

机构信息

Center for Human Genetics, University of Leuven, Belgium.

出版信息

DNA Cell Biol. 1990 Jul-Aug;9(6):461-9. doi: 10.1089/dna.1990.9.461.

Abstract

The polymerase chain reaction (PCR) technique has provided a substantial improvement for the detection and analysis of known genetic polymorphisms. Here, we describe the application of this method for the detection of variable number of tandem repeat (VNTR) sequences. With the use of unique oligonucleotide primers, flanking the repeat sequence, and the thermostable Taq DNA polymerase, the hypervariable regions 3' of the Ha-ras gene, 3' of the apolipoprotein B gene, and 5' to the joining segments of the heavy-chain immunoglobulin gene could be amplified. Alleles up to 2,000 bp could be visualized directly on ethidium bromide-stained agarose gels. Larger alleles were seen only after traditional Southern blot analysis with an internal probe. The value of this new approach for the detection of VNTRs is illustrated in a case of paternity dispute.

摘要

聚合酶链反应(PCR)技术在已知基因多态性的检测和分析方面有了显著改进。在此,我们描述该方法在检测可变数目串联重复序列(VNTR)中的应用。通过使用位于重复序列两侧的独特寡核苷酸引物以及热稳定的Taq DNA聚合酶,可扩增Ha-ras基因3'端的高变区、载脂蛋白B基因3'端以及重链免疫球蛋白基因连接段5'端。长达2000 bp的等位基因可直接在溴化乙锭染色的琼脂糖凝胶上观察到。只有在使用内部探针进行传统的Southern印迹分析后,才能看到更大的等位基因。在一例亲子鉴定纠纷中说明了这种检测VNTR新方法的价值。

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