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“Blind” mapping of genic DNA sequence polymorphisms in Lolium perenne L. by high resolution melting curve analysis

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Abstract

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.).

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Acknowledgements

The authors would like to acknowledge Alistair Hurst from Biotech-IgG A/S, Copenhagen, Denmark, for excellent technical support, Steven Dobrowolski, Idaho Technology, Inc., for inspiring discussions and Johan van Ooijen for helpful advices used for linkage mapping.

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Authors and Affiliations

  1. Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Research Centre Flakkebjerg, Aarhus University, Forsøgsvej 1, 4200, Slagelse, Denmark

    Bruno Studer, Louise Bach Jensen, Alice Fiil & Torben Asp

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  1. Bruno Studer
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  2. Louise Bach Jensen
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  3. Alice Fiil
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  4. Torben Asp
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Correspondence to Bruno Studer.

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Studer, B., Jensen, L.B., Fiil, A. et al. “Blind” mapping of genic DNA sequence polymorphisms in Lolium perenne L. by high resolution melting curve analysis. Mol Breeding 24, 191–199 (2009). https://doi.org/10.1007/s11032-009-9291-x

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  • DOI: https://doi.org/10.1007/s11032-009-9291-x

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