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Complex FISH probes that cover large sections of genomic DNA often contain a high proportion of scattered repetitive sequences that can hinder the visualization of specific signals. Various approaches have been developed to overcome this challenge, depending on the experimental setup and probe quality. A classic strategy for suppressing repetitive sequences is to use unlabeled competitive DNA, such as sheared whole-genome DNA or fractions rich in repetitive elements. In this chapter, we present two protocols: The first describes a rapid method for isolating COT DNA and its application in various FISH experiments, while the second is designed for COT-free FISH with complex probes and uses a special software tool for image enhancement.

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FISH With and Without COT DNA

  • 实验方案

  • 发表日期：

    2026年

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  • Vladimir A. Trifonov,
  • Nadezhda V. Vorobieva,
  • Natalia A. Serdyukova,
  • Willem Rens

摘要

Complex FISH probes that cover large sections of genomic DNA often contain a high proportion of scattered repetitive sequences that can hinder the visualization of specific signals. Various approaches have been developed to overcome this challenge, depending on the experimental setup and probe quality. A classic strategy for suppressing repetitive sequences is to use unlabeled competitive DNA, such as sheared whole-genome DNA or fractions rich in repetitive elements. In this chapter, we present two protocols: The first describes a rapid method for isolating COT DNA and its application in various FISH experiments, while the second is designed for COT-free FISH with complex probes and uses a special software tool for image enhancement.