Quantitative assays for uracil-DNA glycosylase of high sensitivity
- PMID: 3370147
- DOI: 10.1139/o88-021
Quantitative assays for uracil-DNA glycosylase of high sensitivity
Abstract
We have developed a sensitive fluorometric assay using bisulfite deaminated (C----U), covalently-closed circular PM2 DNA as the substrate. We describe a reliable way to prepare this substrate without nicking the PM2 DNA. Methods, which depend on toluenization of the cells, are described for reproducibly and quantitatively assaying uracil-DNA glycosylase. The sensitivity is such that only 200 EL4 mouse thymoma cells or 30,000 Escherichia coli cells are needed for each point in a kinetic assay.
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