Measurement of chromosomal DNA single-strand breaks and replication fork progression rates.
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.
Breslin
Claire
C
Genome Damage and Stability Center, University of Sussex, Brighton, United Kingdom.
Clements
Paula M
PM
El-Khamisy
Sherif F
SF
Petermann
Eva
E
Iles
Natasha
N
Caldecott
Keith W
KW
D000818:Animals; D016466:CHO Cells; D002846:Chromatography, Affinity; D002875:Chromosomes; D020552:Comet Assay; D006224:Cricetinae; D004249:DNA Damage; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000235:genetics*; D004587:Electrophoresis, Agar Gel
D004277:DNA, Single-Stranded