Location: Cereal Crops ResearchTitle: Chromosome painting by GISH and multi-color FISH Author
|Liu, Zhao - North Dakota State University|
|Zhang, Qijun - North Dakota State University|
|Niu, Zhixia - Former ARS Employee|
|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.
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.