Studer, Bruno4; Jensen, Louise Bach1; Fiil, Alice4; Asp, Torben4
1 Molekylær Genetik og Bioteknologi, Faculty of Agricultural Sciences, Aarhus University, Aarhus University2 Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Aarhus University, Aarhus University3 Department of Molecular Biology and Genetics - Afgrødegenetik og Bioteknologi, Department of Molecular Biology and Genetics, Science and Technology, Aarhus University4 Department of Molecular Biology and Genetics - Afgrødegenetik og Bioteknologi, Department of Molecular Biology and Genetics, Science and Technology, Aarhus University
High resolution melting curve analysis (HRM) measures dissociation of double stranded DNA of a PCR product amplified in the presence of a saturating fluorescence dye. Recently, HRM proved successful to genotype DNA sequence polymorphisms such as SSRs and SNPs based on the shape of the melting curves. In this study, HRM was used for simultaneous screening and genotyping of genic DNA sequence polymorphisms identified in the Lolium perenne F2 mapping population VrnA. Melting profiles of PCR products amplified from previously published gene loci and from a novel gene putatively involved in vernalization response successfully discriminated genotypes in absence of allelic sequence information, and allowed to determine allele segregation in VrnA. Here we introduce the concept of "blind" mapping based on HRM as a powerful, fast and cheap method to map any DNA sequence polymorphisms without prior knowledge of allelic sequences in the key grassland species perennial ryegrass (Lolium perenne L.).
Molecular Breeding, 2009, Vol 24, Issue 2, p. 191-199
“Blind” mapping; High resolution melting curve analysis (HRM); Linkage mapping; Lolium perenne L