Measurement of chromosomal DNA single-strand breaks and replication fork progression rates

Abstract

Chromosomal single-strand breaks (SSBs) are the most common lesions arising in cells, but are normally rapidly repaired by multiprotein complexes centered around the scaffold protein, XRCC1. Here, we describe protocols to measure chromosomal SSBs in cells and for recovering and identifying novel components of SSBR complexes in vitro and in vivo. We also describe an assay we employ to measure the rate of replication fork progression in mammalian/vertebrate cells in the presence or absence of DNA damage.