The Random Oligonucleotide-Primed Synthesis Assay for the Quantification of DNA Strand Breaks.

ABSTRACT

DNA strand breaks arise from normal cellular processes such as replication, transcription, and DNA repair as well as spontaneous DNA damage caused by cell metabolic activities. In addition, strand breaks occur due to direct or indirect DNA damage produced by various abiotic and biotic stresses. Strand breaks are among the most problematic DNA lesions because unrepaired strand breaks may lead to cell cycle arrest, gross chromosome rearrangements, or even cell death. Thus, the measurement of the relative number of strand breaks can provide an informative picture of genome stability of a given cell, tissue, or organism. Here, we describe the use of random oligonucleotide-primed synthesis (ROPS) assay for the detection and quantification of the level of strand breaks in tissue samples. The applications of the assay for a quantitative detection of 3'OH, 3'P, or DNA strand breaks at a cleavage site of the deoxyribose residue are discussed.