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3D Multicolor DNA FISH Tool to Study Nuclear Architecture in Human Primary Cells
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Genetics
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JoVE Journal Genetics
3D Multicolor DNA FISH Tool to Study Nuclear Architecture in Human Primary Cells

3D Multicolor DNA FISH Tool to Study Nuclear Architecture in Human Primary Cells

DOI:
10.3791/60712-v

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11:25 min

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January 25, 2020

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, , ,
1Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, INGM

Chapters

  • 00:04Title
  • 00:45DNA Probe Preparation and Nick Translation Labelling
  • 02:15Human Primary T Cell Fixation, Pre-Treatment, and Permeabilization
  • 04:50Human Primary Myoblast Cell Fixation, Pre-Treatment, and Permeabilization
  • 05:433D multicolor DNA FISH
  • 07:053D Multicolor DNA FISH Detection and Analysis
  • 08:52Results: Representative DNA FISH Analysis of Human Primary Cell Nuclear Architecture
  • 10:45Conclusion

Summary

Automatic Translation

Automatic Translation

3D multicolor DNA FISH represents a tool to visualize multiple genomic loci within 3D preserved nuclei, unambiguously defining their reciprocal interactions and localization within the nuclear space at a single cell level. Here, a step by step protocol is described for a wide spectrum of human primary cells.

Tags

Keywords: 3D Multicolor DNA FISHNuclear ArchitectureGenomic LociChromosome CaptureNick TranslationProbe PrecipitationCell FixationPermeabilizationRNAse Treatment

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