FISH with and Without COT1 DNA

  • Vladimir A. TrifonovEmail author
  • Nadezhda V. Vorobieva
  • Natalia A. Serdyukova
  • Willem Rens
Part of the Springer Protocols Handbooks book series (SPH)


Complex FISH probes comprising large spans of genomic DNA always contain a high amount of dispersed repetitive sequences hampering the visualization of specific signals. To overcome this problem, different approaches have been elaborated that depend on experiment type and probe quality. A classical way to suppress repetitive sequences is to use unlabelled competitor DNA (sheared total genomic DNA or repeated sequences enriched DNA fractions). Here we present two protocols—the first one describes a rapid COT DNA isolation and peculiarities of its use in different FISH experiments, and the second is elaborated for COT-free FISH with complex probes and is based on a special software tool for image enhancement.


Repetitive DNA DNA reassociation kinetics Genome composition COT1-DNA 



This work was supported by grant of RSF (№14-14-00275) to VAT and by Welcome Trust grant to WR.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Vladimir A. Trifonov
    • 1
    Email author
  • Nadezhda V. Vorobieva
    • 1
  • Natalia A. Serdyukova
    • 1
  • Willem Rens
    • 2
  1. 1.Institute of Molecular and Cellular Biology SB RASNovosibirskRussian Federation
  2. 2.Department of Veterinary MedicineUniversity of CambridgeCambridgeUnited Kingdom

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