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Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox
JoVE 신문
생화학
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JoVE 신문 생화학
Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

DOI:
10.3791/62378-v

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07:12 min

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July 05, 2021

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, , ,
1Sheffield Institute for Nucleic Acids, Department of Chemistry,University of Sheffield

Chapters

  • 00:04Introduction
  • 00:41Experimental Setup
  • 03:49Preparation Validation
  • 04:24Concentration Bingo
  • 05:41Results: Doubly Labeled Duplex DNA Sample Analysis
  • 06:39Conclusion

Summary

자동 번역

자동 번역

This article provides step-by-step instructions for making fully-corrected accurate FRET measurements on individual, freely diffusing biomolecules using the open-source, inexpensive smfBox, from switch on, through alignment and focusing, to data collection and analysis.

Tags

Single-molecule FRETSmfBoxOpen-sourceBiomolecular SystemsProtein FoldingProtein-DNA InteractionsDNA AllosteryALEXLaser ExcitationFluorescent DyeAlignmentPinhole Positioner

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