+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Single molecule FRET data analysis procedures for FRET efficiency determination: probing the conformations of nucleic acid structures

Single molecule FRET data analysis procedures for FRET efficiency determination: probing the conformations of nucleic acid structures

Methods 64(1): 36-42

Single molecule FRET microscopy is an attractive technique for studying structural dynamics and conformational diversity of nucleic acid structures. Some of its strengths are that it can follow structural changes on a fast time scale and identify conformation distributions arising from dynamic or static population heterogeneity. Here, we give a description of the experiment and data analysis procedures of this method and detail what parameters are needed for FRET efficiency calculation. Using single molecule FRET data obtained on G-quadruplex DNA structures that exhibit large conformation diversity, we illustrate that the shape of the FRET distribution changes depending on what parameters are included in the data analysis procedure.

(PDF same-day service: $19.90)

Accession: 036846686

Download citation: RISBibTeXText

PMID: 23583888

DOI: 10.1016/j.ymeth.2013.04.001

Related references

BOBA FRET: bootstrap-based analysis of single-molecule FRET data. Plos One 8(12): E84157-E84157, 2014

Maximizing information content of single-molecule FRET experiments: multi-color FRET and FRET combined with force or torque. Chemical Society Reviews 43(4): 1007-1013, 2014

Integrated magnetic tweezers and single-molecule FRET for investigating the mechanical properties of nucleic acid. Methods 105: 16-25, 2018

Future Perspective of Single-Molecule FRET Biosensors and Intravital FRET Microscopy. Biophysical Journal 111(6): 1103-1111, 2017

Probing Protein Interactions Using GFP and FRET * Stage 2: Cell Preparation for FLIM-FRET Analysis. Csh Protocols 2006(1): -, 2006

Manipulating protein conformations by single-molecule AFM-FRET nanoscopy. Acs Nano 6(2): 1221-1229, 2012

Accurate single molecule FRET efficiency determination for surface immobilized DNA using maximum likelihood calculated lifetimes. Journal of Physical Chemistry. B 111(11): 2986-2990, 2007

LacI-DNA-IPTG loops: equilibria among conformations by single-molecule FRET. Journal of Physical Chemistry. B 117(16): 4713-4722, 2013

Single-Molecule FRET Reveals Three Conformations for the TLS Domain of Brome Mosaic Virus Genome. Biophysical Journal 109(12): 2625-2636, 2016

Molecular dynamics simulation of configurational ensembles compatible with experimental FRET efficiency data through a restraint on instantaneous FRET efficiencies. Journal of Computational Chemistry 35(32): 2319-2332, 2015

Intravital FRET: comprehending life at single-molecule level. Focus on "A practical method for monitoring FRET-based biosensors in living animals using two-photon microscopy". American Journal of Physiology. Cell Physiology 309(11): C722-C723, 2016

Probing the dynamics of restriction endonuclease NgoMIV-DNA interaction by single-molecule FRET. Biopolymers 107(12): -, 2017

Probing the Conformational Landscape of DNA Polymerases Using Diffusion-Based Single-Molecule FRET. Methods in Enzymology 581: 353-378, 2016

Structural dynamics of nucleic acids by single-molecule FRET. Methods in Cell Biology 113: 1-37, 2013

Ensemble and single-molecule detected time-resolved FRET methods in studies of protein conformations and dynamics. Methods in Molecular Biology 1076: 113-169, 2014