S'identifier

University of Waikato

2 ARTICLES PUBLISHED IN JoVE

Genetics

Measuring Microbial Mutation Rates with the Fluctuation Assay

Rok Krašovec¹, Huw Richards^2,3, Guillaume Gomez³, Danna R. Gifford¹, Adrien Mazoyer⁴, Christopher G. Knight³

¹School of Biological Sciences, The University of Manchester, ²School of Science, University of Waikato, ³School of Natural Sciences, The University of Manchester, ⁴Department of Mathematics, Université du Québec à Montréal

Here, a protocol is presented to perform a fluctuation assay and estimate microbial mutation rate using phenotypic markers. This protocol will enable researchers to assay mutations in diverse microbes and environments, determining how genotype and ecological context affect spontaneous mutation rates.

Biochemistry

Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes

Ronja Stelzer¹, Elizabeth Rzoska-Smith¹, Sigurd Gundesø², Ulli Rothweiler², Adele Williamson¹

¹University of Waikato, ²ArcticZymes Technologies ASA

Here, a protocol for assaying nucleic acid metabolizing enzymes is presented, using examples of ligase, nuclease, and polymerase enzymes. The assay utilizes fluorescently labeled and unlabeled oligonucleotides that can be combined to form duplexes mimicking RNA and/or DNA damages or pathway intermediates, allowing for the characterization of enzyme behavior.