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Detection of single nucleotide mutations in wheat using single strand conformation polymorphism gels

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Abstract

We show that single strand conformation polymorphism (SSCP) analysis, using the mutation detection enhancement (MDETM) matrix, is efficient at detecting sequence polymorphisms in PCR amplicons. Four independent wheat genomic fragments were amplified from two contrasting templates and sequenced. The allelic fragments were differentiated at 1–6 single nucleotide positions, but MDE-SSCP was able to unequivocally distinguish each allelic pair. The approach is therefore considered a powerful way of identifying single nucleotide polymorphisms (SNPs) without extensive amplicon sequencing.

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Abbreviations

MDE:

mutation detection enhancement

SNP:

single nucleotide polymorphism

SSCP:

single strand conformation polymorphism

TBE:

tris borate ethylene-diaminetetraacetic acid

TEMED:

tetrametylethylenediamine

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Authors and Affiliations

  1. Departamento de Gen’etica e Biotecnologia, ICETA-Universidade de Tr’as-os-Montes e Alto Douro, Apartado 202, 5001, Vila Real, Portugal

    P. Martins-Lopes

  2. John Innes Centre, Norwich Research Park, Colney, NR4 7UH, Norwich, U.K.

    H. Zhang & R. Koebner

Authors
  1. P. Martins-Lopes
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  2. H. Zhang
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  3. R. Koebner
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Martins-Lopes, P., Zhang, H. & Koebner, R. Detection of single nucleotide mutations in wheat using single strand conformation polymorphism gels. Plant Mol Biol Rep 19, 159–162 (2001). https://doi.org/10.1007/BF02772158

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  • DOI: https://doi.org/10.1007/BF02772158

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