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

Photochem Photobiol. 2002 Aug;76(2):123-6. doi: 10.1562/0031-8655(2002)076<0123:dvoaso>2.0.co;2.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry*
  • DNA Damage*
  • Microscopy, Fluorescence
  • Photochemistry

Substances

  • DNA