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Live-Cell Imaging of Transcriptional Activity at DNA Double-Strand Breaks
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Biology
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JoVE Journal Biology
Live-Cell Imaging of Transcriptional Activity at DNA Double-Strand Breaks

Live-Cell Imaging of Transcriptional Activity at DNA Double-Strand Breaks

DOI:
10.3791/62968-v

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09:07 min

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September 20, 2021

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, , ,
1Instituto de Medicina Molecular João Lobo Antunes,Faculdade de Medicina da Universidade de Lisboa

Chapters

  • 00:04Introduction
  • 00:53Preparation and Transfection of Cells for Live-Cell Microscopy
  • 02:09Experimental Setup
  • 04:05Image Acquisition
  • 06:04Microscopy Calibration Measurements and Analysis
  • 06:51Results: Analysis of Transcription Detection at Sites of DNA Double-Strand Breaks
  • 08:12Conclusion

Summary

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This protocol presents a new reporter gene system and the experimental setup to detect transcription at DNA double-strand breaks with single-molecule sensitivity.

Tags

Keywords: Live-cell ImagingTranscriptional ActivityDNA Double-strand BreaksTranscription DetectionSingle-cell AnalysisDNA Damage ResponseTranscription-replication CrosstalkDoxycycline InductionCalibration MeasurementsMicrocentrifuge Tube PreparationTransfectionGlass Bottom DishMicroscopy Observation

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