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

MPG-Autoren
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Borodina,  Tatiana A.
Technology Development(Alexey Soldatov), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Lehrach,  Hans
Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Soldatov,  Aleksey V.
Technology Development(Alexey Soldatov), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Zitation

Borodina, T. A., Lehrach, H., & Soldatov, A. V. (2004). Ligation detection reaction-TaqMan procedure for single nucleotide polymorphism detection on genomic DNA. Analytical Biochemistry, 333, 309-319. doi:10.1016/j.ab.2004.05.032.


Zitierlink: https://hdl.handle.net/11858/00-001M-0000-0010-87A2-9
Zusammenfassung
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.