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Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
Journal JoVE
Biologie
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Journal JoVE Biologie
Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

DOI:
10.3791/3255-v

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07:18 min

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October 27, 2011

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1Department of Molecular Oncology, Weatherall Institute of Molecular Medicine,University of Oxford , 2Institute of Genetics and Biotechnology, Faculty of Biology,University of Warsaw

Chapitres

  • 00:05Titre
  • 01:21In vivo Labeling DT40 Cells
  • 02:04Cell Lysis and DNA Spreading
  • 02:53Immunofluorescence Staining
  • 04:31Image Acquisition and Data Analysis
  • 05:24Replication Structures
  • 06:54Conclusion

Summary

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Traduction automatique

DT40, a model vertebrate genetic system, provides a powerful tool to analyze protein function. Here we describe a simple method that allows qualitative analysis of parameters that influence DNA synthesis during the S-phase in DT40 cells at the single molecule level.

Tags

DNA ReplicationVisualizationVertebrate Model SystemDT40DNA Fiber TechniqueReplication Fork StabilityGenome DuplicationEndogenous DNA DamageExogenous DNA DamageGenomic InstabilityTumor DevelopmentFluorescence MicroscopyAvian B-cell Line DT40Protein FunctionReverse GeneticsDNA Replication And Genome StabilityReplication Fork DynamicsDNA Synthesis RateQualitative AnalysisDNA Fiber TechniqueHalogenated Nucleotides

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