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A slot-blot hybridisation method for screening somatic hybrids

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Abstract

A slot-blot method is described that provides rapid and reliable identification of somatic hybrids. Biotin-labelled probes of defined length were prepared from genomic DNA, and calibrated in conjunction with target DNA to allow for differing amounts of biotin incorporation and hybridisation efficiency. The presence of probe DNA was visualised by chemiluminescence detection. Probes of 1 to 5 kbp in length gave optimal discrimination among target DNAs from different potato species. The advantages of this method over existing ones are described.

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Abbreviations

PEG:

polyethylene glycol

RAPD:

random amplified polymorphic DNA

CTAB:

cetyltrimethylammoniumbromide

AMPPD:

3-(2′-spiroadamantane)-4-methoxy-4-(3′-phosphoryloxy)phenyl-1,2-dioxetane

GISH:

genomicin-situ hybridisation

References

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Authors and Affiliations

  1. Molecular Biology and Diagnostics Section, National Institute of Agricultural Boany, Huntingdon Road, CB3OLE, Cambridge, UK

    D. Matthews, K. Harding, M. J. Wilkinson & S. Millam

Authors
  1. D. Matthews
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  2. K. Harding
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  3. M. J. Wilkinson
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  4. S. Millam
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Corresponding author

Correspondence to D. Matthews.

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Matthews, D., Harding, K., Wilkinson, M.J. et al. A slot-blot hybridisation method for screening somatic hybrids. Plant Mol Biol Rep 15, 62–70 (1997). https://doi.org/10.1007/BF02772113

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  • DOI: https://doi.org/10.1007/BF02772113

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