Ligation detection reaction-TaqMan procedure for single nucleotide polymorphism detection on genomic DNA

被引:20
作者
Borodina, TA [1 ]
Lehrach, H [1 ]
Soldatov, AV [1 ]
机构
[1] Max Planck Inst Mol Genet, D-14195 Berlin, Germany
关键词
SNP genotyping; TaqMan; mutation; detection;
D O I
10.1016/j.ab.2004.05.032
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:309 / 319
页数:11
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