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Chromosome Painting by GISH and Multicolor FISH

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Plant Cytogenetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1429))

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.

Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.

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Acknowledgment

The authors thank Drs. Bikram S. Gill and Bernd Friebe (Kansas State University, Manhattan, KS, USA) for providing the original protocols. We also thank Drs. Xueyong Zhang (Chinese Academy of Agricultural Sciences, Beijing, China), Qi Zheng (Chinese Academy of Sciences, Beijing, China), Lili Qi (USDA-ARS, Fargo, ND, USA), and Jiming Jiang (University of Wisconsin, Madison, WI, USA) for the instruction and discussion of the procedure. Dr. Jiming Jiang kindly provided the plasmids of 5S rDNA and 45S rDNA. This work was supported by the USDA-ARS CRIS Project No. 3060-520-037-00D. USDA is an equal opportunity provider and employer.

Author information

Authors and Affiliations

  1. USDA-ARS, Cereal Crops Research Unit, Northern Crop Science Laboratory, 1605 Albrecht Blvd. North, Fargo, ND, 58102, USA

    Steven S. Xu & Zhixia Niu

  2. Department of Plant Sciences, North Dakota State University, Fargo, ND, 58108, USA

    Zhao Liu, Qijun Zhang & Xiwen Cai

  3. USDA-ARS, Sunflower and Plant Biology Research Unit, Northern Crop Science Laboratory, Fargo, ND, 58102, USA

    Chao-Chien Jan

Authors
  1. Steven S. Xu
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  2. Zhao Liu
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  3. Qijun Zhang
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  4. Zhixia Niu
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  5. Chao-Chien Jan
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  6. Xiwen Cai
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Corresponding author

Correspondence to Steven S. Xu .

Editor information

Editors and Affiliations

  1. University of Minnesota, St. Paul, Minnesota, USA

    Shahryar F. Kianian

  2. University of Minnesota, St. Paul, Minnesota, USA

    Penny M. Avoles Kianian

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Xu, S.S., Liu, Z., Zhang, Q., Niu, Z., Jan, CC., Cai, X. (2016). Chromosome Painting by GISH and Multicolor FISH. In: Kianian, S., Kianian, P. (eds) Plant Cytogenetics. Methods in Molecular Biology, vol 1429. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3622-9_2

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  • DOI: https://doi.org/10.1007/978-1-4939-3622-9_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3620-5

  • Online ISBN: 978-1-4939-3622-9

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