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Flow Sorting Plant Chromosomes

  • Jan VránaEmail author
  • Petr Cápal
  • Jarmila Číhalíková
  • Marie Kubaláková
  • Jaroslav Doležel
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1429)

Abstract

Nuclear genomes of many important plant species are tremendously complicated to map and sequence. The ability to isolate single chromosomes, which represent small units of nuclear genome, is priceless in many areas of plant research including cytogenetics, genomics, and proteomics. Flow cytometry is the only technique which can provide large quantities of pure chromosome fractions suitable for downstream applications including physical mapping, preparation of chromosome-specific BAC libraries, sequencing, and optical mapping. Here, we describe step-by-step procedure of preparation of liquid suspensions of intact mitotic metaphase chromosomes and their flow cytometric analysis and sorting.

Key words

Cell cycle synchronization Chromosome isolation Cytogenetic stocks FISH FISHIS Flow cytometry and sorting Metaphase accumulation Plants 

Notes

Acknowledgment

This work was supported by grant LO1204 from the National Program of Sustainability I. We also thank Ms. Zdeňka Dubská and Ms. Romana Šperková for excellent technical assistance.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jan Vrána
    • 1
    Email author
  • Petr Cápal
    • 1
  • Jarmila Číhalíková
    • 1
  • Marie Kubaláková
    • 1
  • Jaroslav Doležel
    • 1
  1. 1.Centre of the Region Haná for Biotechnological and Agricultural ResearchInstitute of Experimental BotanyOlomoucCzech Republic

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