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Published on: September 25, 2018
Ligation detection reaction-TaqMan procedure for single nucleotide polymorphism detection on genomic DNA
Tatiana A Borodina1, Hans Lehrach, Aleksey V Soldatov
1Max Planck Institut für Molekulare Genetik, 14195 Berlin-Dahlem, Germany.
Abstract:
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.
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