SYBR Green I: Fluorescence Properties and Interaction with DNA

被引：228

作者：

Dragan, A. I. ^{[1
]}

Pavlovic, R. ^{[1
]}

McGivney, J. B. ^{[3
]}

Casas-Finet, J. R. ^{[3
]}

Bishop, E. S. ^{[4
]}

Strouse, R. J. ^{[3
]}

Schenerman, M. A. ^{[3
]}

Geddes, C. D. ^{[1
,2
]}

机构：

[1] Univ Maryland Baltimore Cty, Inst Fluorescence, Baltimore, MD 21202 USA

[2] Univ Maryland Baltimore Cty, Dept Chem & Biochem, Baltimore, MD 21202 USA

[3] Medimmune Inc, Gaithersburg, MD 20878 USA

[4] Cygnus Technol, Southport, NC 28461 USA

来源：

JOURNAL OF FLUORESCENCE | 2012年 / 22卷 / 04期

关键词：

Syber Green; Picogreen; Metal-enhanced fluorescence; Fluorescence; Correlation spectroscopy; DNA; Intercalating dyes; DOUBLE-STRANDED DNA; NUCLEIC-ACIDS; PHOTOPHYSICAL PROPERTIES; ETHIDIUM-BROMIDE; HOECHST; 33258; BINDING; PICOGREEN; QUANTITATION; ASSAY; STAIN;

D O I：

10.1007/s10895-012-1059-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In this study, we have investigated the fluorescence properties of SYBR Green I (SG) dye and its interaction with double-stranded DNA (dsDNA). SG/dsDNA complexes were studied using various spectroscopic techniques, including fluorescence resonance energy transfer and time-resolved fluorescence techniques. It is shown that SG quenching in the free state has an intrinsic intramolecular origin; thus, the observed > 1,000-fold SG fluorescence enhancement in complex with DNA can be explained by a dampening of its intra-molecular motions. Analysis of the obtained SG/DNA binding isotherms in solutions of different ionic strength and of SG/DNA association in the presence of a DNA minor groove binder, Hoechst 33258, revealed multiple modes of interaction of SG inner groups with DNA. In addition to interaction within the DNA minor groove, both intercalation between base pairs and stabilization of the electrostatic SG/DNA complex contributed to increased SG affinity to double-stranded DNA. We show that both fluorescence and the excited state lifetime of SG dramatically increase in viscous solvents, demonstrating an approximate 200-fold enhancement in 100 % glycerol, compared to water, which also makes SG a prospective fluorescent viscosity probe. A proposed structural model of the SG/DNA complex is compared and discussed with results recently reported for the closely related PicoGreen chromophore.

引用

页码：1189 / 1199

页数：11

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