Direct visualization of abasic sites on a single DNA molecule using fluorescence microscopy.

Photochem Photobiol

Plant Resources Laboratory, Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, 1233 Watanuki-machi, Takasaki, Gunma 370-1292, Japan.

Published: August 2002

A new method was developed to allow direct visualization of damaged sites on individual DNA molecules. Fluorescence in situ hybridization on extended DNA molecules was modified to detect a single abasic site. Abasic sites were specifically labeled with a biotinylated aldehyde-reactive probe and fluorochrome-conjugated streptavidin. The light emitted by a single fluorochrome-DNA complex was calibrated. The number of abasic sites on the DNA molecule was estimated by counting each fluorochrome-DNA complex. The present study directly visualized and characterized the abasic sites of single DNA molecules.

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http://dx.doi.org/10.1562/0031-8655(2002)076<0123:dvoaso>2.0.co;2DOI Listing

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