A simple procedure for the analysis of single nucleotide polymorphisms facilitates map-based cloning in Arabidopsis
A simple procedure for the analysis of single nucleotide polymorphisms facilitates map-based cloning in Arabidopsis
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DOI：
10.1104/pp.124.4.1483
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发表时间：
2000-12-01
期刊：
影响因子：
7.4
通讯作者：
Ausubel, FM
中科院分区：
文献类型：
--
作者：
Drenkard, E;Richter, BG;Ausubel, FM
我们开发了一种改良的等位基因特异性 PCR 程序来测定单核苷酸多态性 (SNP)，并使用该程序（称为 SNAP，用于单核苷酸扩增多态性）生成 62 个拟南芥作图标记。 SNAP 引物在一个等位基因（特异性等位基因）3' 端的三个核苷酸内含有单碱基对错配，此外还与非特异性等位基因存在 3' 错配。使用名为 SNAPER 的计算机程序来促进引物设计，使特异性和非特异性 SNP 等位基因的扩增产物数量产生至少 1,000 倍的差异。由于 SNAP 标记可以通过标准琼脂糖凝胶上的电泳轻松测定，并且由于拟南芥哥伦比亚和兰茨伯格直立生态型之间存在超过 25,000 个 SNP 的公共数据库，因此 SNAP 方法极大地促进了由突变表型定义的拟南芥基因的基于图谱的克隆。
We developed a modified allele-specific PCR procedure for assaying single nucleotide polymorphisms (SNPs) and used the procedure (called SNAP for single-nucleotide amplified polymorphisms) to generate 62 Arabidopsis mapping markers. SNAP primers contain a single base pair mismatch within three nucleotides from the 3' end of one allele (the specific allele) and in addition have a 3' mismatch with the nonspecific allele. A computer program called SNAPER was used to facilitate the design of primers that generate at least a 1,000-fold difference in the quantity of the amplification products from the specific and nonspecific SNP alleles. Because SNAP markers can be readily assayed by electrophoresis on standard agarose gels and because a public database of over 25,000 SNPs is available between the Arabidopsis Columbia and Landsberg erectra ecotypes, the SNAP method greatly facilitates the map-based cloning of Arabidopsis genes defined by a mutant phenotype.