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

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)44-50
Number of pages7
JournalAnalytical Biochemistry
Volume441
Issue number1
DOIs
StatePublished - Jan 1 2013

Keywords

  • DNA
  • Förster resonance energy transfer
  • Single molecule

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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