Laser Flash Photolysis Studies of Dna‐complexed Ethidium Bromide

Atherton, Stephen J.; Beaumont, Paul C.

doi:10.1111/j.1751-1097.1986.tb03570.x

Cited by 13 publications

(14 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…10 At higher nucleotide-to-dye ratios, where negligible amounts of free Eb exist, the decay becomes m ono-exponential, and the large lifetime reaches a m aximum value between 21 and 24 ns, according to different authors. 2,8,9,11 This lifetime is then associated with the excited state lifetime of the intercalated Eb. W hen the ionic strength is reduced below 0.05 M, an intermediate-lived component between the free and intercalated values is consistently obtained 2 and it has been associated with the lifetime of Eb binding at the secondary sites.…”

Section: Introductionmentioning

confidence: 99%

Photophysical and Direct Determination of Binding Constants of Ethidium Bromide Complexed to E. Coli DNA

et al. 2002

View full text Add to dashboard Cite

High-precision time-correlated single-photon-counting data were obtained to achieve the resolution of the fluorescence decay time components from DNA-Ethidium bromide (Eb) buffered solutions at low (0.05 M) and high (0.4 M) NaCl concentrations. Tri-exponential functions are the best models to describe the decay kinetics in all solutions, independently of the NaCl concentration. The fluorescence nanosecond graphs were recorded intercalating polarizers into the light path to check for anisotropy artifacts. The results suggest that the three exponentials from fluorescence decay graphs depict three true lifetimes, which, in general, are dependent on the [DNA]/[Eb] ratio and can be assigned to free Eb and bound Eb at the low- and high-affinity DNA sites. With the normalized weighting coefficients from tri-exponential fitting of nanosecond decay graphs, recorded at the excitation wavelength of the isosbestic point, it has been possible to calculate the values of binding constants and site numbers at the two salt concentrations by means of the neighbor exclusion model (McGhee–von Hipple expression).

show abstract

Section: Introductionmentioning

confidence: 99%

Photophysical and Direct Determination of Binding Constants of Ethidium Bromide Complexed to E. Coli DNA

et al. 2002

View full text Add to dashboard Cite

show abstract

“…The similar long lifetime (22.2 ns) for ethidium intercalated between DNA bases has been attributed to a reduction in the rate of excited state proton transfer to solvent molecules [29]. Lifetimes of this magnitude are not observed when ethidium is dissolved in a wide range of hydrophobic solvents [2].…”

Section: Ethidiummentioning

confidence: 67%

Spatial Relationships between Drug Binding Sites on the Surface of the Acetylcholine Receptor

Johnson¹

1986

View full text Add to dashboard Cite

“…The most notable advantage of FP-DBPs for staining DNA is that they do not mediate DNA photocleavage, which makes them ideal staining reagents for imaging of DNA molecules over extended time periods. As shown in Figure 3 ( see Supplementary Movie 1a), DNA staining with organic dyes commonly leads to DNA breakage because illuminated fluorophores continually cycle through excitation and emission, which greatly enhances photochemical reactions causing DNA photocleavage ( 17 , 29 ). Accordingly, DNA fluorescence studies involving organic dyes require careful illumination dosages and copious addition of anti-bleaching agents (e.g.…”

Section: Resultsmentioning

confidence: 99%

DNA binding fluorescent proteins for the direct visualization of large DNA molecules

Lee¹,

Oh²,

Lee³

et al. 2015

Nucleic Acids Res

View full text Add to dashboard Cite

Fluorescent proteins that also bind DNA molecules are useful reagents for a broad range of biological applications because they can be optically localized and tracked within cells, or provide versatile labels for in vitro experiments. We report a novel design for a fluorescent, DNA-binding protein (FP-DBP) that completely ‘paints’ entire DNA molecules, whereby sequence-independent DNA binding is accomplished by linking a fluorescent protein to two small peptides (KWKWKKA) using lysine for binding to the DNA phosphates, and tryptophan for intercalating between DNA bases. Importantly, this ubiquitous binding motif enables fluorescent proteins (Kd = 14.7 μM) to confluently stain DNA molecules and such binding is reversible via pH shifts. These proteins offer useful robust advantages for single DNA molecule studies: lack of fluorophore mediated photocleavage and staining that does not perturb polymer contour lengths. Accordingly, we demonstrate confluent staining of naked DNA molecules presented within microfluidic devices, or localized within live bacterial cells.

show abstract

Laser Flash Photolysis Studies of Dna‐complexed Ethidium Bromide

Cited by 13 publications

References 8 publications

Photophysical and Direct Determination of Binding Constants of Ethidium Bromide Complexed to E. Coli DNA

Photophysical and Direct Determination of Binding Constants of Ethidium Bromide Complexed to E. Coli DNA

Spatial Relationships between Drug Binding Sites on the Surface of the Acetylcholine Receptor

DNA binding fluorescent proteins for the direct visualization of large DNA molecules

Contact Info

Product

Resources

About