Ligation detection reaction-TaqMan procedure for single nucleotide polymorphism detection on genomic DNA
Ligation detection reaction-TaqMan procedure for single nucleotide polymorphism detection on genomic DNA
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DOI：
10.1016/j.ab.2004.05.032
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发表时间：
2004-10-15
影响因子：
2.9
通讯作者：
Soldatov, AV
中科院分区：
文献类型：
--
作者：
Borodina, TA;Lehrach, H;Soldatov, AV
在这篇文章中，我们描述了一种基因分型方法，适用于个人和多重单核苷酸多态性（SNP）分析，连接检测反应（LDR）直接进行基因组DNA的基础上。在连接过程中，SNP基因座的双等位基因状态转化为连接的检测寡核苷酸的双标记物状态。然后用通用荧光探针通过5 '-核酸酶测定（TaqMan）确定标记物的状态。LDR-TaqMan方法成功应用于拟南芥30个SNP位点的基因分型。该技术具有成本效益，不需要特定于位置的优化，需要最少的操作，并且具有非常好的自动化潜力。(C)2004年爱思唯尔公司All rights reserved.
In this article, we describe a genotyping approach applicable to both individual and multiplexed single nucleotide polymorphism (SNP) analysis, based on a ligation detection reaction (LDR) performed directly on genomic DNA. During the ligation, the biallelic state of the SNP locus is converted into a bimarker state of ligated detector oligonucleotides. The state of the markers is then determined by a 5'-nuclease assay (TaqMan) with universal fluorescent probes. The LDR-TaqMan method was successfully applied for the genotyping of 30 SNP loci of Arabidopsis thaliana. The technology is cost-effective, needs no locus-specific optimization, requires minimal manipulations, and has very good potential for automation. (C) 2004 Elsevier Inc. All rights reserved.