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  • Title: Preparation of samples for comparative studies of plant chromosomes using in situ hybridization methods.
    Author: Walling JG, Pires JC, Jackson SA.
    Journal: Methods Enzymol; 2005; 395():443-60. PubMed ID: 15865979.
    Abstract:
    The development of fluorescence in situ hybridization (FISH) has led to the advancement of chromosome studies not only for physical mapping and genome analyses but also as a tool for evolutionary studies. Isolated repetitive DNA sequences have been useful cytological markers, but large-insert genomic libraries (e.g., bacterial artificial chromosomes [BACs], yeast artificial chromosomes [YACs], and cosmids) are being increasingly used to serve as probes for large segments of DNA across related genomes. Although FISH is usually applied to metaphase chromosomes, fiber FISH, a variation of FISH using extended DNA fibers, is now used to measure loci at the resolution of a few kilobases to compare orthologous genome segments across related genomes. A generalized set of protocols for chromosome preparation, FISH, and fiber FISH are presented; however, it is often necessary to experiment with techniques for different plant taxa for successful molecular cytogenetic studies.
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