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[长片段精确PCR(LA PCR)]

[Long and accurate PCR (LA PCR)].

作者信息

Mukai H, Nakagawa T

机构信息

Advanced Biomedical Center, Takara Shuzo Co., LTD.

出版信息

Nihon Rinsho. 1996 Apr;54(4):917-22.

PMID:8920650
Abstract

PCR (Polymerase Chain Reaction) has become a standard and essential tool used in disciplines such as molecular genetics, genome mapping and sequencing, and forensics. It has proven to be easier, quicker, and less costly than traditional techniques of molecular cloning. While its use in these fielde has already contributed to great advancements and revolutionary changes, current PCR using Taq DNA polymerase is usually limited to amplification up to 5kb. Recently, "long" PCR conditions have been identified which allow the amplification of DNA templates up to 40kb with higher fidelity. In this report, we describe the amplification of longer human genomic fragments (up to 27kb) using LA PCR technology.

摘要

聚合酶链反应(PCR)已成为分子遗传学、基因组图谱绘制与测序以及法医学等学科中使用的标准且必不可少的工具。事实证明,它比传统的分子克隆技术更简便、快捷且成本更低。虽然它在这些领域的应用已经带来了巨大的进步和革命性的变化,但目前使用Taq DNA聚合酶的PCR通常仅限于扩增长度达5kb的片段。最近,已确定了“长片段”PCR条件,可实现对长达40kb的DNA模板进行更高保真度的扩增。在本报告中,我们描述了使用LA PCR技术扩增更长的人类基因组片段(长达27kb)的情况。

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