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

作者：Kelliher Michael T; Piraino Mark S; Gemoules Madeline E; Southern Cathrine A^*

来源：Analytical Biochemistry, 2013, 441(1): 44-50.

DOI：10.1016/j.ab.2013.06.009

• Forster resonance energy transfer
• Single molecule
• DNA

Ensemble Forster 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.

• 出版日期2013-10-1