1. Determination of Mitotic Recombination Rates by Fluctuation Analysis in Saccharomyces cerevisiae
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The study of recombination in Saccharomyces cerevisiae benefits from the availability of assay systems that select for recombinants, allowing the study of spontaneous events that
represent natural assaults on the genome. However, the rarity of such spontaneous recombination requires selection of events
that occur over many generations in a cell culture, and the number of recombinants increases exponentially following a recombination
event. To avoid inflation of the average number of recombinants by jackpots arising from an event early in a culture, the
distribution of the number of recombinants in independent cultures (fluctuation analysis) must be used to estimate the mean
number of recombination events. Here we describe two statistical analyses (method of the median and the method of p0) to estimate the true mean of the number of events to be used to calculate the recombination rate. The use of confidence
intervals to depict the error in such experiments is also discussed. The application of these methods is illustrated using
the intron-based inverted repeat recombination reporter system developed in our lab to study the regulation of homeologous
recombination.
Book Title: Genetic Recombination: Reviews and Protocols
Series: Methods in Molecular Biology | Volume: 262 | Pub. Date: Jan-09-2004 | Page Range: 3-12 | DOI: 10.1385/1-59259-761-0:003
Subject: Genetics/Genomics
Key Words: fluctuation analysis - method of the median - confidence intervals - spontaneous recombination - mutation rate - inverted repeats - intron-based recombination assay - homeologous recombination
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