Panhandle PCR.
Panhandle PCR.
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狭长地带 PCR。
DOI：
10.1101/gr.4.5.s195
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Jones,DH
中科院分区：
文献类型：
--
作者：
Jones,DH
PCR允许体外高度特异性的DNA扩增。PCR通过从序列的每个末端延伸引物而发生，因此需要了解这些末端。(1)已经开发了几种方法用于PCR扩增已知序列一端侧翼的未知DNA。（z-zS）这些方法将PCR的应用扩展到DNA的检索，其中仅DNA序列的一端是已知的，使得人们可以使用PCR沿未表征的DNA片段”行走”沿着，而无需筛选文库中的重叠克隆。除了柄状PCR之外，现有的方法都不允许对位于已知位点侧翼的> 3.0kb的人基因组DNA进行高度特异性的PCR扩增。本文描述了一种用于柄状PCR的方案，该方案允许直接从大量人类基因组DNA中高度特异性地扩增约<4.4kb的DNA，该DNA位于已知位点的侧翼。
PCR permits highly specific DNA amplification in vitro. PCR occurs by primer extension from each end of a sequence and therefore requires knowledge of those ends.(1) Several methods have been developed for PCR amplification of unknown DNA that flanks one end of a known sequence.(z-zS) These methods extend the application of PCR to the retrieval of DNA where only one end of the DNA sequence is known, so that one can use PCR to" walk" along an uncharacterized stretch of DNA without screening a library for overlapping clones. None of the existing methods, apart from panhandle PCR, has permired the highly specific PCR amplification of> 3.0 kb of human genomic DNA that flanks a known site. This paper describes a protocol for panhandle PCR that permits the highly specific PCR amplification of~< 4.4 kb of DNA that flanks a known site directly from bulk human genomic DNA.