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. 1994 Jan;2(1):15-20.
doi: 10.1007/BF01539448.

Sensitivity enhancement of fluorescence in situ hybridization on plant chromosomes

W Busch  1 , R Martin, R G Herrmann
Affiliations

Sensitivity enhancement of fluorescence in situ hybridization on plant chromosomes

W Busch et al. Chromosome Res. 1994 Jan.

Abstract

An improved in 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 for in 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 low-copy 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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