Ratiometric analysis of single-molecule fluorescence resonance energy transfer using logical combinations of threshold criteria: A study of 12-mer DNA

被引：50

作者：

Ying, LM ^{[1
]}

Wallace, MI ^{[1
]}

Balasubramanian, S ^{[1
]}

Klenerman, D ^{[1
]}

机构：

[1] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England

来源：

JOURNAL OF PHYSICAL CHEMISTRY B | 2000年 / 104卷 / 21期

关键词：

D O I：

10.1021/jp993914k

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

Single-molecule fluorescence resonance energy transfer (FRET) combined with bulk fluorescence lifetimes, anisotropy, and spectra have been used to study a donor-acceptor labeled model DNA system (Cy5-5'-ACCTGCCGACGC-3'-TMR). A general ratiometric analysis method using independent donor and acceptor thresholding has been developed. Use of two logical combinations of thresholding criteria provides more information than either method alone, revealing heterogeneity within this system. Conditions yielding similar bulk fluorescence spectra can be readily distinguished by this single-molecule method. Fluorescence lifetimes and anisotropy measurements also suggest nonnegligible fluorophore-DNA interaction.

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收藏

页码：5171 / 5178

页数：8

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