Mutation accumulation (MA) experiments are widely used in evolutionary biology for studying the effects, rates and properties of new spontaneous mutations. Despite this, they are laborious to execute, typically taking months to years to a acquire sufficient mutations to measure mutation rates and spectra. Further, typical tools such as agar plates and repetitive streaking are poor representations of real environmental living conditions. Methods for determining MA in conditions that better represent the environment of an organism are needed.
Researchers at the Biodesign Institute of Arizona State University have developed a novel liquid-phase mutation accumulation experiment, which more accurately reflects an organism’s environment. Starting with an ancestor line, serial dilution is employed across multiple concentrations followed by culturing on a 96-deepwell plate, selecting a turbid well from the lowest serial dilution for subsequent dilution rounds and repeating. This experiment allows for a wider variety of mutations to occur in a shorter amount of time so that sufficient mutations can occur fur measuring mutation rates and spectra. Liquid handling laboratory equipment can be utilized to help automate the process and minimize human derived errors.
With its promise of automation and scalability, this technology holds great promise in reducing the human capital requirements of measuring mutation rates and advancing evolutionary biology.
Potential Applications
- Liquid mutation accumulation experiments
- Measure mutagenic effect of toxins or other variables (could be used by industry or government to understand safety of chemicals or materials)
Benefits and Advantages
- Can be automated either fully or partially to speed up the process and minimize human errors
- Allows for a wider variety of mutations in a shorter amount of time
- Exhibits slightly more cell divisions per day than extant protocols for measuring MA
- Could potentially be connected to plate readers to measure OD600 and get proxy measurement of cell growth
- Slightly modified mutation spectrum can be obtained
- Reduces human capital requirements in measuring mutation rates
- Immune to various human biases in colony selection
- Fitness decline was not significantly different between plate and liquid lines
- May protect some error-prone bacteria from some oxidative damage
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