Drenkard E, Richter B G, Rozen S, Stutius L M, Angell N A, Mindrinos M, Cho R J, Oefner P J, Davis R W, Ausubel F M
Department of Genetics, Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Plant Physiol. 2000 Dec;124(4):1483-92. doi: 10.1104/pp.124.4.1483.
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 erecta ecotypes, the SNAP method greatly facilitates the map-based cloning of Arabidopsis genes defined by a mutant phenotype.
我们开发了一种改良的等位基因特异性PCR程序用于检测单核苷酸多态性（SNP），并使用该程序（称为单核苷酸扩增多态性的SNAP）生成了62个拟南芥定位标记。SNAP引物在一个等位基因（特异性等位基因）3'端的三个核苷酸内包含一个单碱基对错配，此外与非特异性等位基因存在3'端错配。使用一个名为SNAPER的计算机程序来辅助设计引物，使得特异性和非特异性SNP等位基因的扩增产物数量至少相差1000倍。由于SNAP标记可以很容易地通过在标准琼脂糖凝胶上进行电泳来检测，并且在拟南芥哥伦比亚生态型和直立型生态型之间有一个超过25000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图位克隆。
