title: Preparation of samples for comparative studies of plant chromosomes using in situ hybridization methods.
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.
authors.affiliation: Department of Agronomy, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: Jason G
authors.initials: JG
authors.lastname: Walling
authors.forename: J Chris
authors.initials: JC
authors.lastname: Pires
authors.forename: Scott A
authors.initials: SA
authors.lastname: Jackson
mesh_terms: D005075:Biological Evolution; D002458:Cell Fractionation / Q000379:methods; D032461:Chromosomes, Plant / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008540:Meiosis / Q000235:genetics; D008938:Mitosis / Q000235:genetics; D015336:Molecular Probe Techniques; D009691:Nucleic Acid Denaturation; D018517:Plant Roots / Q000235:genetics; D010944:Plants / Q000235:genetics*
chemical_list: D018744:DNA, Plant