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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Vol. 76 • No. 2