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Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage
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Biochemistry
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JoVE Journal Biochemistry
Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage

Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage

English

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1,704 Views

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09:53 min

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February 07, 2021

DOI:

10.3791/62112-v

DOI:
10.3791/62112-v

•

09:53 min
February 07, 2021

•

2 Views
, , , ,
1Department of Chemistry,Tufts University, 2Department of Physics,Wesleyan University, 3Wyss Institute at Harvard University

Summary

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Using quantum-dot-labeled DNA and total internal reflection fluorescence microscopy, we can investigate the reaction mechanism of restriction endonucleases while using unlabeled protein. This single-molecule technique allows for massively multiplexed observation of individual protein-DNA interactions, and data can be pooled to generate well-populated dwell-time distributions.

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Cite this Article

Etson, C. M., Todorov, P., Shatery Nejad, N., Shrestha, N., Walt, D. R. Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage. J. Vis. Exp. (168), e62112, doi:10.3791/62112 (2021).

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