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Large-scale mapping using high-molecular-weight DNA fromP. vulgaris

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Abstract

The application of megabase methods is essential for positional cloning of genes for which no products have been characterized. We have set about the isolation of genes conferring resistance to pathogens in the french beanPhaseolus vulgaris. We describe here the isolation of high-molecular-weight DNA, pulse-field gel electrophoresis and large-scale mapping of thePal2 locus inP. vulgaris. We describe the construction of such a map by embedding DNA isolated from leaf nuclei in agarose plugs, restricting it with rare cutters and subjecting the DNA to pulse-field gel electrophoresis.

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Abbreviations

PFGE:

pulse-field gel electrophoresis

BSA:

bovine serum albumin

PAL:

phenylalanine ammonia lyase

CHEF:

contour clamped homogeneous electric field

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Authors and Affiliations

  1. Département de Biologie Moléculaire Végétale, URA CNRS 1128, Bâtiment 430, Université Paris Sud, 91405, Orsay, France

    Francine Creusot, Mireille Sevignac & Michel Dron

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  1. Francine Creusot
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  2. Mireille Sevignac
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  3. Michel Dron
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Creusot, F., Sevignac, M. & Dron, M. Large-scale mapping using high-molecular-weight DNA fromP. vulgaris . Plant Mol Biol Rep 10, 207–218 (1992). https://doi.org/10.1007/BF02668348

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