TY - JOUR
T1 - A comparison of Förster resonance energy transfer analysis approaches for Nanodrop fluorometry
AU - Kelliher, Michael T.
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
Y1 - 2013
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
KW - Single molecule
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U2 - 10.1016/j.ab.2013.06.009
DO - 10.1016/j.ab.2013.06.009
M3 - Article
C2 - 23811157
AN - SCOPUS:84881125620
VL - 441
SP - 44
EP - 50
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -