Quantifying Double-Strand Breaks and Clustered Damages in DNA by Single-Molecule Laser Fluorescence Sizing

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Elena M. Filippova (Creator)

Denise C. Monteleone (Creator)

Stephen R. Quake (Creator)

Betsy M. Sutherland (Creator)

John C. Sutherland (Creator)

John G. Trunk (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: Fluorescence from a single DNA molecule passing through a laser beam is proportional to the size (contour length) of the molecule and molecules of different sizes can be counted with equal efficiencies. Single-molecule fluorescence can thus determine the average length of the molecules in a sample and hence the frequency of double-strand breaks induced by various treatments. Ionizing radiation-induced frank double-strand breaks can thus be quantified by single-molecule sizing. Moreover multiple classes of clustered damages involving damaged bases and abasic sites alone or in combination with frank single-strand breaks can be quantified by converting them to double-strand breaks by chemical or enzymatic treatments. For a given size range of DNA molecules single-molecule sizing is as or more sensitive than gel electrophoresis and requires several orders-of-magnitude less DNA to determine damage levels. Originally published Biophysical Journal Vol. 84 No. 2 Feb 2003

Additional Information

Publication

Other

Biophysical Journal. 84:2(February 2003) p. 1281-1290.

Language: English

Date: 2011

Keywords

Laser Fluorescence Sizing, DNA, double-strand breaks, cluster damage

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