Abstract
Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon that has been repurposed as a biophysical tool to measure nanometer distances. With FRET by DNA eXchange, or FRET X, many points of interest (POIs) in a single object can be probed, overcoming a major limitation of conventional single-molecule FRET. In FRET X, short fluorescently labeled DNA imager strands specifically and transiently bind their complementary docking strands on a target molecule, such that at most a single FRET pair is formed at each point in time and multiple POIs on a single molecule can be readily probed. Here, we describe the sample preparation, image acquisition, and data analysis for structural analysis of DNA nanostructures with FRET X.
Original language | English |
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Title of host publication | Methods in Molecular Biology |
Publisher | Humana Press Inc. |
Pages | 203-213 |
Number of pages | 11 |
DOIs | |
State | Published - 2024 |
Publication series
Name | Methods in Molecular Biology |
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Volume | 2694 |
ISSN (Print) | 1064-3745 |
ISSN (Electronic) | 1940-6029 |
Bibliographical note
Publisher Copyright:© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.
Keywords
- DNA nanotechnology
- FRET X
- Single-molecule FRET
- Single-molecule multiplexing
- Structural biology