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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Cereal Crops Research » Research » Publications at this Location » Publication #314187

Research Project: Genetic Improvement of Durum and Spring Wheat for Quality and Resistance to Diseases and Pests

Location: Cereal Crops Research

Title: Chromosome painting by GISH and multi-color FISH

Author
item Xu, Steven
item Liu, Zhao - North Dakota State University
item Zhang, Qijun - North Dakota State University
item Niu, Zhixia - Former ARS Employee
item Jan, Chao-chien
item Cai, Xiwen - North Dakota State University

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: 8/10/2015
Publication Date: 4/1/2016
Citation: Xu, S.S., Liu, Z., Zhang, Q., Niu, Z., Jan, C., Cai, X. 2016. Chromosome Painting by GISH and Multicolor FISH. In: Kianian, S.F., Kianian, P.M.A., Editors. Plant Cytogenetics: Methods and Protocols. Methods in Molecular Biology. New York: Springer. p. 7-21.

Interpretive Summary:

Technical 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 multi-color 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. Multi-color 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.