In vitro monitoring of base excision repair in saccharomyces cerevisiae

Abstract

Base excision repair (BER) is an important mechanism to maintain genomic stability. Here we offer a set of protocols to quantitatively analyze BER capacity in whole cell-free yeast extracts. Cell-free yeast extracts were obtained by a French press procedure and repair capacities were measured by using oligonucleotide substrates. Repair products were separated by polyacrylamide gel electrophoresis and detected by autoradiography. These set of methods allow the analysis of different kinds of base damage and of individual mechanistic steps within BER. We used these protocols to investigate a new role of the DNA double strand break repair protein XRS1 in BER (1). © 2012 Springer Science+Business Media New York.