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.
Hirose
Tamaki
T
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
Ohtani
Toshio
T
Muramatsu
Hiroshi
H
Tanaka
Atsushi
A
D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D008856:Microscopy, Fluorescence; D010777:Photochemistry
D004247:DNA