Direct visualization of abasic sites on a single DNA molecule using fluorescence microscopy.
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.
Plant Resources Laboratory, Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, 1233 Watanuki-machi, Takasaki, Gunma 370-1292, Japan. thirose@taka.jaeri.go.jp
Tamaki
T
Hirose
Toshio
T
Ohtani
Hiroshi
H
Muramatsu
Atsushi
A
Tanaka
D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D008856:Microscopy, Fluorescence; D010777:Photochemistry
D004247:DNA