Sensitivity enhancement of fluorescencein situ hybridization on plant chromosomes |
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Authors: | W. Busch R. Martin R. G. Herrmann |
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Affiliation: | (1) Botanisches Institut, Ludwig-Maximilians-Universität, Menzinger Str. 67, 80638 München, Germany |
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Abstract: | An improvedin situ hybridization procedure is presented, based on synchronization of root meristems of barley and wheat, enzymatic digestion, a protoplast drop technique, and the use of the fluorescent dye Cy3. The combination of these approaches resulted in a significant increase of well-spread metaphases suitable forin situ hybridization as compared to squash preparations, and to a significantly enhanced number and intensity of hybridization signals as demonstrated for a B-hordein-specific lowcopy probe of barley. In the case of Cy3 all metaphases displayed a signal, more than 60% of them on both chromatids of each gene-bearing chromosome. |
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Keywords: | B-hordein cell cycle synchronization drop spread technique fluorescencein situ hybridization |
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