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Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
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Biochimica
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JoVE Journal Biochimica
Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

DOI:
10.3791/59073-v

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06:24 min

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February 13, 2019

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, , ,
1Cell Biology Center, Institute of Innovative Research,Tokyo Institute of Technology

Capitoli

  • 00:04Titolo
  • 00:52DNA Strand Pairing and Displacement Assays
  • 02:57Estimate Maximum FRET Efficiency of the Pairing and Displacement Assays
  • 03:58Results: Real-time Measurement of Rad51-Driven DNA Strand Exchange
  • 06:06Conclusion

Summary

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Fluorescence resonance energy transfer-based real-time observation systems of the DNA strand exchange reaction mediated by Rad51 were developed. Using the protocols presented here, we are able to detect the formation of reaction intermediates and their conversion into products, while also analyzing the enzymatic kinetics of the reaction.

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Keywords: DNA Strand ExchangeRad51Real-time ObservationRecombination ProteinsDNA Pairing AssayFluorescence SpectroscopyFRET

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