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Fluorescence In Situ Hybridization on DNA Halo Preparations to Reveal Whole Chromosomes, Telomeres and Gene Loci
JoVE 杂志
生物学
This content is Free Access.
JoVE 杂志 生物学
Fluorescence In Situ Hybridization on DNA Halo Preparations to Reveal Whole Chromosomes, Telomeres and Gene Loci

Fluorescence In Situ Hybridization on DNA Halo Preparations to Reveal Whole Chromosomes, Telomeres and Gene Loci

DOI:
10.3791/62017-v

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09:07 min

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March 04, 2021

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, , ,
1Laboratory of Nuclear and Genomic Health, Centre for Genome Engineering and Maintenance, Division of Biosciences, Department of Life Sciences, College of Health, Medicine and Life Sciences,Brunel University London, 2Biosciences, Department of Clinical, Pharmaceutical and Biological Science, School of Life and Medical Sciences,University of Hertfordshire

Chapters

  • 00:04Introduction
  • 00:46Slide Preparation and Cell Culture
  • 02:08DNA Halo Preparation
  • 03:22Two‐Dimensional Fluorescence In Situ Hybridization
  • 06:47Results: DNA Halos for High‐Resolution Analysis of Genomic Interactions with the Nucleoskeleton
  • 08:41Conclusion

Summary

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Combining DNA halo preparations with fluorescence in situ hybridization enables high-resolution analysis of genomic interactions with the nucleoskeleton. Attached genome leads to hybridized fluorescent signals located within the residual extracted nuclei, whereas non-attached genome is in the halo of DNA surrounding the residual nuclei.

Tags

Keywords: DNA HaloFluorescence In Situ Hybridization (FISH)NucleoskeletonSingle Cell AssaySpatial OrganizationFunctional OrganizationGenomeSenescent CellsCancer CellsProgeria CellsModel OrganismsCytoskeleton BufferDNA Halo BufferExtraction BufferEthanol Series

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