A comparison of Förster resonance energy transfer analysis approaches for Nanodrop fluorometry

Michael T. Kelliher; Mark Stephens Piraino; Madeline E. Gemoules; Cathrine A. Southern

doi:10.1016/j.ab.2013.06.009

A comparison of Förster resonance energy transfer analysis approaches for Nanodrop fluorometry

Michael T. Kelliher, Mark Stephens Piraino, Madeline E. Gemoules, Cathrine A. Southern^*

^*Corresponding author for this work

Mathematics

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

Ensemble Förster resonance energy transfer (FRET) results can be analyzed in a variety of ways. Due to experimental artifacts, the results obtained from different analysis approaches are not always the same. To determine the optimal analysis approach to use for Nanodrop fluorometry, we have performed both ensemble and single-molecule FRET studies on oligomers of double-stranded DNA. We compared the single- molecule FRET results with those obtained using various ensemble FRET analysis approaches. This comparison shows that for Nanodrop fluorometry, analyzing the increase of the acceptor fluorescence is less likely to introduce errors in estimates of FRET efficiencies compared with analyzing the fluorescence intensity of the donor in the absence and presence of the acceptor.

Original language	English (US)
Pages (from-to)	44-50
Number of pages	7
Journal	Analytical Biochemistry
Volume	441
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2013.06.009
State	Published - 2013

Funding

This work was funded by the Camille and Henry Dreyfus Start-up and Supplemental Award Programs , the DePaul University Department of Chemistry , and the DePaul University Research Council Competitive Research Grant Program . The authors thank Caitlin Karver and Richard Niedziela for carefully reviewing this manuscript.

Keywords

DNA
Förster resonance energy transfer
Single molecule

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.ab.2013.06.009

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title = "A comparison of F{\"o}rster resonance energy transfer analysis approaches for Nanodrop fluorometry",

abstract = "Ensemble F{\"o}rster resonance energy transfer (FRET) results can be analyzed in a variety of ways. Due to experimental artifacts, the results obtained from different analysis approaches are not always the same. To determine the optimal analysis approach to use for Nanodrop fluorometry, we have performed both ensemble and single-molecule FRET studies on oligomers of double-stranded DNA. We compared the single- molecule FRET results with those obtained using various ensemble FRET analysis approaches. This comparison shows that for Nanodrop fluorometry, analyzing the increase of the acceptor fluorescence is less likely to introduce errors in estimates of FRET efficiencies compared with analyzing the fluorescence intensity of the donor in the absence and presence of the acceptor.",

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note = "Funding Information: This work was funded by the Camille and Henry Dreyfus Start-up and Supplemental Award Programs , the DePaul University Department of Chemistry , and the DePaul University Research Council Competitive Research Grant Program . The authors thank Caitlin Karver and Richard Niedziela for carefully reviewing this manuscript.",

year = "2013",

doi = "10.1016/j.ab.2013.06.009",

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AU - Piraino, Mark Stephens

AU - Gemoules, Madeline E.

AU - Southern, Cathrine A.

N1 - Funding Information: This work was funded by the Camille and Henry Dreyfus Start-up and Supplemental Award Programs , the DePaul University Department of Chemistry , and the DePaul University Research Council Competitive Research Grant Program . The authors thank Caitlin Karver and Richard Niedziela for carefully reviewing this manuscript.

PY - 2013

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N2 - Ensemble Förster resonance energy transfer (FRET) results can be analyzed in a variety of ways. Due to experimental artifacts, the results obtained from different analysis approaches are not always the same. To determine the optimal analysis approach to use for Nanodrop fluorometry, we have performed both ensemble and single-molecule FRET studies on oligomers of double-stranded DNA. We compared the single- molecule FRET results with those obtained using various ensemble FRET analysis approaches. This comparison shows that for Nanodrop fluorometry, analyzing the increase of the acceptor fluorescence is less likely to introduce errors in estimates of FRET efficiencies compared with analyzing the fluorescence intensity of the donor in the absence and presence of the acceptor.

AB - Ensemble Förster resonance energy transfer (FRET) results can be analyzed in a variety of ways. Due to experimental artifacts, the results obtained from different analysis approaches are not always the same. To determine the optimal analysis approach to use for Nanodrop fluorometry, we have performed both ensemble and single-molecule FRET studies on oligomers of double-stranded DNA. We compared the single- molecule FRET results with those obtained using various ensemble FRET analysis approaches. This comparison shows that for Nanodrop fluorometry, analyzing the increase of the acceptor fluorescence is less likely to introduce errors in estimates of FRET efficiencies compared with analyzing the fluorescence intensity of the donor in the absence and presence of the acceptor.

KW - DNA

KW - Förster resonance energy transfer

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A comparison of Förster resonance energy transfer analysis approaches for Nanodrop fluorometry

Abstract

Funding

Keywords

ASJC Scopus subject areas

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