Standardized Protocols from the Caenorhabditis Intervention Testing Program 2013-2016: Conditions and Assays used for Quantifying the Development, Fertility and Lifespan of Hermaphroditic Caenorhabditis Strains

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Abstract

The Caenorhabditis Intervention Testing Program \(CITP) seeks to identify compounds that robustly improve lifespan and/or attenuate age-dependent declines in health across a genetically diverse set of Caenorhabditis species and strains. A core mission of the CITP is to collect data that will inform on the reproducibility of our experimental results. To achieve this, multiple replicates of each experiment are performed by each of three groups at separate laboratory sites. In an effort to limit non-biological confounding factors that may in uence the reproducibility of these results, we sought to minimize differences in experimental conditions across the individual laboratories. This led to the development and utilization of a set of standardized protocols that describe speci c methods for nematode maintenance and culture as well as detailed experimental procedures. All laboratories of the CITP are tasked with strict adherence to protocols. Here we provide several protocols that describe the CITP standard operating practices for bacterial cultures, media preparation and strain handing as well as assays for quantifying the development rate, fertility and lifespan of hermaphroditic Caenorhabditis strains. Individual components of these methods were not tested for their effects on outcomes and are not suggested to be best practices. However our consideration of the methodological variability available for even these simple assays does highlight the potential for differences in methodology to in uence experimental reproducibility.

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