Detection of abasic sites on individual DNA molecules using atomic force microscopy

H. B. Sun, L. Qian, H. Yokota

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

We have developed an atomic force microscopy-based method for detecting abasic sites (AP sites) on individual DNA molecules. By using uracil and uracil DNA glycosylase, we first prepared a 250-bp DNA template consisting of two AP sites at specific locations. We then detected the AP sites by marking them with biotinylated aldehyde-reactive probes and monomeric avidin. We demonstrate here that (i) the location of monomeric avidin bound on a single DNA molecule was detectable by atomic force microscopy; (ii) the observed location of avidin was in good agreement to the predicted AP sites at a few nanometer resolution; and (iii) by end-labeling the 5′-terminus of one DNA strand, the AP sites were determined without directional ambiguity. The technique described here will provide a sensitive way of locating AP sites and contribute to screen DNA damages from individual molecules.

Original languageEnglish (US)
Pages (from-to)2229-2232
Number of pages4
JournalAnalytical Chemistry
Volume73
Issue number10
DOIs
StatePublished - May 15 2001
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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