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使用热稳定DNA聚合酶进行引物引导的DNA酶促扩增。

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.

作者信息

Saiki R K, Gelfand D H, Stoffel S, Scharf S J, Higuchi R, Horn G T, Mullis K B, Erlich H A

机构信息

Cetus Corporation, Department of Human Genetics, Emeryville, CA 94608.

出版信息

Science. 1988 Jan 29;239(4839):487-91. doi: 10.1126/science.2448875.

Abstract

A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction. The enzyme, isolated from Thermus aquaticus, greatly simplifies the procedure and, by enabling the amplification reaction to be performed at higher temperatures, significantly improves the specificity, yield, sensitivity, and length of products that can be amplified. Single-copy genomic sequences were amplified by a factor of more than 10 million with very high specificity, and DNA segments up to 2000 base pairs were readily amplified. In addition, the method was used to amplify and detect a target DNA molecule present only once in a sample of 10(5) cells.

摘要

一种热稳定的DNA聚合酶被用于体外DNA扩增程序,即聚合酶链反应。这种从嗜热水生栖热菌中分离出来的酶极大地简化了该程序,并且通过使扩增反应能够在更高温度下进行,显著提高了可扩增产物的特异性、产量、灵敏度和长度。单拷贝基因组序列以非常高的特异性扩增了超过1000万倍,并且长达2000个碱基对的DNA片段很容易被扩增。此外,该方法被用于扩增和检测在10⁵个细胞的样本中仅出现一次的目标DNA分子。

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