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GrainGenes Reference Report: GNM-44-523

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Reference	GNM-44-523
Title	Application of denaturing high-performance liquid chromatography for mapping of single nucleotide polymorphisms in barley (Hordeum vulgare L.)
Journal	Genome
Year	2001
Volume	44
Pages	523-528
Author	Kota R Wolf M Michalek W Graner A
Abstract	Summary: Recent advances in DNA sequence analysis and the establishment of high-throughput assays have provided the framework for large-scale discovery and analysis of DNA sequence variation. In this context, single nucleotide polymorphisms (SNPs) are of particular interest. To initiate a systematic approach to develop an SNP map of barley (Hordeum vulgare L.), we have employed denaturing high-performance liquid chromatography (DHPLC) to analyse segregating SNP patterns in a doubled-haploid (DH) mapping population. To this end, SNPs between the parental genotypes were identified using a direct sequencing approach. Once a SNP was established between the parents, the optimal melting temperature of the PCR fragment containing the SNP was predicted for its analysis by DHPLC. Following the detection of the optimal temperature, the DH lines were analysed for the presence of either of the alleles. To test the utility of the analysis, data from previously mapped RFLP markers from which these SNPs were derived were compared. Results from these experiments indicate that DHPLC can be efficiently employed in analysing SNPs on a high-throughput scale
Keyword	[ Hide all but 1 of 47 ] alleles assay barley canada chromatography denaturing high performance liquid chromatography detection dh lines direct sequencing dna sequence doubled-haploid doubled-haploid lines establishment framework genetic mapping genome genotype high performance liquid chromatography hordeum hordeum vulgare hybridization large-scale line liquid chromatography map mapping marker melting microsatellite molecular markers mutation nucleotide oligonucleotides pattern pcr polymorphism population rflp rflp marker scale sequence sequence analysis sequence variation sequencing single snp snp map

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