title: Chromosome Painting by GISH and Multicolor FISH.
abstract: Fluorescent in situ hybridization (FISH) is a powerful cytogenetic technique for identifying chromosomes and mapping specific genes and DNA sequences on individual chromosomes. Genomic in situ hybridization (GISH) and multicolor FISH (mc-FISH) represent two special types of FISH techniques. Both GISH and mc-FISH experiments have general steps and features of FISH, including chromosome preparation, probe labeling, blocking DNA preparation, target-probe DNA hybridization, post-hybridization washes, and hybridization signal detection. Specifically, GISH uses total genomic DNA from two species as probe and blocking DNA, respectively, and it can differentiate chromosomes from different genomes. The mc-FISH takes advantage of simultaneous hybridization of several DNA probes labeled by different fluorochromes to different targets on the same chromosome sample. Hybridization signals from different probes are detected using different fluorescence filter sets. Multicolor FISH can provide more structural details for target chromosomes than single-color FISH. In this chapter, we present the general experimental procedures for these two techniques with specific details in the critical steps we have modified in our laboratories.
authors: USDA-ARS, Cereal Crops Research Unit, Northern Crop Science Laboratory, 1605 Albrecht Blvd. North, Fargo, ND, 58102, USA. steven.xu@ars.usda.gov.
authors: Steven S
authors: SS
authors: Xu
authors: Department of Plant Sciences, North Dakota State University, Fargo, ND, 58108, USA.
authors: Zhao
authors: Z
authors: Liu
authors: Department of Plant Sciences, North Dakota State University, Fargo, ND, 58108, USA.
authors: Qijun
authors: Q
authors: Zhang
authors: USDA-ARS, Cereal Crops Research Unit, Northern Crop Science Laboratory, 1605 Albrecht Blvd. North, Fargo, ND, 58102, USA.
authors: Zhixia
authors: Z
authors: Niu
authors: USDA-ARS, Sunflower and Plant Biology Research Unit, Northern Crop Science Laboratory, Fargo, ND, 58102, USA.
authors: Chao-Chien
authors: CC
authors: Jan
authors: Department of Plant Sciences, North Dakota State University, Fargo, ND, 58108, USA.
authors: Xiwen
authors: X
authors: Cai
mesh_terms: D020223:Chromosome Painting / Q000379:methods*; D032461:Chromosomes, Plant; D055028:Comparative Genomic Hybridization / Q000379:methods*; D015342:DNA Probes; D018745:Genome, Plant; D006368:Helianthus / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D014908:Triticum / Q000235:genetics*
chemical_list: D015342:DNA Probes
keywords: Blocking DNA; DNA probe; Fluorescent in situ hybridization; Genomic in situ hybridization; Molecular cytogenetics