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Generation of Paint Probes from Flow-Sorted and Microdissected Chromosomes

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Fluorescence In Situ Hybridization (FISH)

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

FISH with whole chromosome or region-specific painting probes made from either flow-sorted or microdissected chromosomes has revolutionized cytogenetics. Generation of paints from flow-sorted chromosomes relies on the use of an expensive and sophisticated fluorescence-activated cell sorter and suspensions of freshly prepared chromosomes. Preparation of paints from microdissected materials requires an inverted microscope with appropriate micromanipulators and metaphase chromosome spreads on coverslips. Painting probes made from flow-sorted chromosomes generally have better chromosomal coverage and can be used in a wide range of applications between distantly related species, while region-specific probes from microdissection enable higher-resolution analyses restricted to comparative painting between closely related species. Here we provide detailed protocols on generation probes from both flow-sorted and microdissected chromosomes.

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Acknowledgments

While writing this protocol, VT was supported by Budget Projects 0310-2014-0003, 0310-2014-0008, 0310-2014-0009, and DFG. FY, BLN, and NPC are supported by the Wellcome Trust (Grant number WT098051).

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Authors and Affiliations

  1. Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SA, UK

    Fengtang Yang, Bee Ling Ng & Nigel P. Carter

  2. Institute of Molecular and Cellular Biology, Russian Academy of Sciences, 630090, Novosibirsk, Russia

    Vladimir Trifonov

  3. Jena University Hospital, Institute of Human Genetics, Friedrich Schiller University, Kollegiengasse 10, 07743, Jena, Germany

    Nadezda Kosyakova

Authors
  1. Fengtang Yang
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  2. Vladimir Trifonov
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  3. Bee Ling Ng
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  4. Nadezda Kosyakova
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  5. Nigel P. Carter
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Corresponding author

Correspondence to Fengtang Yang .

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Editors and Affiliations

  1. Institut für Humangenetik, Jena, Germany

    Thomas Liehr

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© 2017 Springer-Verlag Berlin Heidelberg

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Yang, F., Trifonov, V., Ng, B.L., Kosyakova, N., Carter, N.P. (2017). Generation of Paint Probes from Flow-Sorted and Microdissected Chromosomes. In: Liehr, T. (eds) Fluorescence In Situ Hybridization (FISH). Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-52959-1_6

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  • DOI: https://doi.org/10.1007/978-3-662-52959-1_6

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  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-52957-7

  • Online ISBN: 978-3-662-52959-1

  • eBook Packages: Springer Protocols

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