Sequence mapping by electronic PCR
Sequence mapping by electronic PCR
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DOI：
10.1101/gr.7.5.541
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发表时间：
1997-05-01
期刊：
影响因子：
7
通讯作者：
Schuler, GD
中科院分区：
文献类型：
--
作者：
Schuler, GD
高度特异和灵敏的聚合酶链式反应为序列标记位点(STSS)提供了基础，STSS是一种独特的标志物，已被广泛用于构建人类基因组的遗传图谱和物理图谱。电子聚合酶链式反应(e-聚合酶链式反应)是指通过寻找与聚合酶链式反应引物紧密匹配、具有正确顺序、方向和间距的子序列来恢复DNA序列中这些独特的位点的过程，这些子序列可以可信地启动具有正确分子量的聚合酶链式反应产物的扩增。开发了一种软件工具，以提供这一搜索策略的有效实施，并允许进行现代基因组分析所需的那种集体搜索。进行了一些样本搜索，以证明可能影响获得匹配的可能性的一些因素。对一个大型序列数据库记录的分析揭示了几个微卫星和基于基因的标记的存在，并使人们能够计算出它们之间的准确碱基对距离。这个例子展示了电子聚合酶链式反应如何用于将不断增长的基因组序列数据与现有图谱相结合，揭示不同图谱上先前存在的标记之间的关系，以及如何将遗传距离与物理距离相关联。
The highly specific and sensitive PCR provides the basis for sequence-tagged sites (STSs), unique landmarks that have been used widely in the construction of genetic and physical maps of the human genome. Electronic PCR (e-PCR) refers to the process of recovering these unique sites in DNA sequences by searching for subsequences that closely match the PCR primers and have the correct order, orientation, and spacing that they could plausibly prime the amplification of a PCR product of the correct molecular weight. A software tool was developed to provide an efficient implementation of this search strategy and allow the sort of en masse searching that is required for modern genome analysis. Some sample searches were performed to demonstrate a number of factors that can affect the likelihood of obtaining a match. Analysis of one large sequence database record revealed the presence of several microsatellite and gene-based markers and allowed the exact base-pair distances among them to be calculated. This example provides a demonstration of how e-PCR can be used to integrate the growing body of genomic sequence data with existing maps, reveal relationships among markers that existed previously on different maps, and correlate genetic distances with physical distances.