Associated anisotropy decays of Ethidium Bromide interacting with DNA

Rahul Chib, Sangram Limbaji Raut, Sarika Sabnis, Preeti Singhal, Zygmunt Gryczynski, Ignacy Gryczynski

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

Ethidium Bromide (EB) is a commonly used dye in a deoxyribonucleic acid (DNA) study. Upon an intercalation, this dye significantly increases its brightness and fluorescence lifetime. In this report we have studied the time resolved fluorescence properties of EB existing simultaneously in free and DNAbound forms in the solution. Fluorescence intensity decays were fitted globally to a double exponential model with lifetimes corresponding to free (1.6 ns) and bound (22 ns) forms, and molar fractions were determined for all used solutions. Anisotropy decays displayed characteristic time dependence with an initial rapid decline followed by recovery and slow decay. The shortlived fraction associated with free EB molecules decreases faster than longlived fraction associated with EB bound to DNA. Consequently, contribution from fast rotation leads to initial rapid decay in anisotropy. On the other hand bound fraction, due to slow rotation helps recover anisotropy in time. This effect of associated anisotropy decays in systems such as EB free/EB-DNA is clearly visible in a wide range of concentrations, and should be taken into account in polarization assays and biomolecule dynamics studies.

Original languageEnglish
Article number015003
JournalMethods and Applications in Fluorescence
Volume2
Issue number1
DOIs
StatePublished - 1 Mar 2014

Fingerprint

Ethidium
bromides
Anisotropy
DNA
deoxyribonucleic acid
anisotropy
Fluorescence
decay
Dyes
fluorescence
Biomolecules
Coloring Agents
Intercalation
dyes
Luminance
Assays
life (durability)
Polarization
Recovery
intercalation

Keywords

  • Associated anisotropy decays
  • DNA fluorescence
  • Ethidium bromide

Cite this

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title = "Associated anisotropy decays of Ethidium Bromide interacting with DNA",
abstract = "Ethidium Bromide (EB) is a commonly used dye in a deoxyribonucleic acid (DNA) study. Upon an intercalation, this dye significantly increases its brightness and fluorescence lifetime. In this report we have studied the time resolved fluorescence properties of EB existing simultaneously in free and DNAbound forms in the solution. Fluorescence intensity decays were fitted globally to a double exponential model with lifetimes corresponding to free (1.6 ns) and bound (22 ns) forms, and molar fractions were determined for all used solutions. Anisotropy decays displayed characteristic time dependence with an initial rapid decline followed by recovery and slow decay. The shortlived fraction associated with free EB molecules decreases faster than longlived fraction associated with EB bound to DNA. Consequently, contribution from fast rotation leads to initial rapid decay in anisotropy. On the other hand bound fraction, due to slow rotation helps recover anisotropy in time. This effect of associated anisotropy decays in systems such as EB free/EB-DNA is clearly visible in a wide range of concentrations, and should be taken into account in polarization assays and biomolecule dynamics studies.",
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year = "2014",
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Associated anisotropy decays of Ethidium Bromide interacting with DNA. / Chib, Rahul; Raut, Sangram Limbaji; Sabnis, Sarika; Singhal, Preeti; Gryczynski, Zygmunt; Gryczynski, Ignacy.

In: Methods and Applications in Fluorescence, Vol. 2, No. 1, 015003, 01.03.2014.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Associated anisotropy decays of Ethidium Bromide interacting with DNA

AU - Chib, Rahul

AU - Raut, Sangram Limbaji

AU - Sabnis, Sarika

AU - Singhal, Preeti

AU - Gryczynski, Zygmunt

AU - Gryczynski, Ignacy

PY - 2014/3/1

Y1 - 2014/3/1

N2 - Ethidium Bromide (EB) is a commonly used dye in a deoxyribonucleic acid (DNA) study. Upon an intercalation, this dye significantly increases its brightness and fluorescence lifetime. In this report we have studied the time resolved fluorescence properties of EB existing simultaneously in free and DNAbound forms in the solution. Fluorescence intensity decays were fitted globally to a double exponential model with lifetimes corresponding to free (1.6 ns) and bound (22 ns) forms, and molar fractions were determined for all used solutions. Anisotropy decays displayed characteristic time dependence with an initial rapid decline followed by recovery and slow decay. The shortlived fraction associated with free EB molecules decreases faster than longlived fraction associated with EB bound to DNA. Consequently, contribution from fast rotation leads to initial rapid decay in anisotropy. On the other hand bound fraction, due to slow rotation helps recover anisotropy in time. This effect of associated anisotropy decays in systems such as EB free/EB-DNA is clearly visible in a wide range of concentrations, and should be taken into account in polarization assays and biomolecule dynamics studies.

AB - Ethidium Bromide (EB) is a commonly used dye in a deoxyribonucleic acid (DNA) study. Upon an intercalation, this dye significantly increases its brightness and fluorescence lifetime. In this report we have studied the time resolved fluorescence properties of EB existing simultaneously in free and DNAbound forms in the solution. Fluorescence intensity decays were fitted globally to a double exponential model with lifetimes corresponding to free (1.6 ns) and bound (22 ns) forms, and molar fractions were determined for all used solutions. Anisotropy decays displayed characteristic time dependence with an initial rapid decline followed by recovery and slow decay. The shortlived fraction associated with free EB molecules decreases faster than longlived fraction associated with EB bound to DNA. Consequently, contribution from fast rotation leads to initial rapid decay in anisotropy. On the other hand bound fraction, due to slow rotation helps recover anisotropy in time. This effect of associated anisotropy decays in systems such as EB free/EB-DNA is clearly visible in a wide range of concentrations, and should be taken into account in polarization assays and biomolecule dynamics studies.

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