A protocol for chemical mutagenesis in Strongyloides ratti

•We present an improved protocol for chemical mutagenesis of Strongyloides ratti.•Relatively high genetic variability complicates the identification of mutations.•Further inbreeding of S. ratti is desirable and tolerated by the organism. Genetic analysis using experimentally induced mutations has been a most valuable tool in the analysis of various organisms. However, genetic analysis of endoparasitic organisms tends to be difficult because of the limited accessibility of the sexually reproducing adults, which are normally located within the host. Nematodes of the genera Strogyloides and Parastrongyloides represent an exception to this because they can form facultative free-living sexually reproducing generations in between parasitic generations. Here we present a protocol for the chemical mutagenesis of Strongyloides ratti. Further we evaluate the feasibility of identifying the induced mutations by whole genome re-sequencing.