High-throughput gene mapping in Caenorhabditis elegans

• Published in: Genome research Vol. 12; no. 7; pp. 1100 - 1105
• Main Authors: Swan, Kathryn A, Curtis, Damian E, McKusick, Kathleen B, Voinov, Alexander V, Mapa, Felipa A, Cancilla, Michael R
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory Press 01.07.2002
• Subjects: Animals; Caenorhabditis elegans - genetics; Chromosome Mapping - methods; Gene Order - genetics; Genes, Helminth - genetics; Methods; Molecular Sequence Data; Physical Chromosome Mapping; Polymorphism, Single Nucleotide - genetics
• ISSN: 1088-9051; 1549-5469
• DOI: 10.1101/gr.208902

Positional cloning of mutations in model genetic systems is a powerful method for the identification of targets of medical and agricultural importance. To facilitate the high-throughput mapping of mutations in Caenorhabditis elegans, we have identified a further 9602 putative new single nucleotide polymorphisms (SNPs) between two C. elegans strains, Bristol N2 and the Hawaiian mapping strain CB4856, by sequencing inserts from a CB4856 genomic DNA library and using an informatics pipeline to compare sequences with the canonical N2 genomic sequence. When combined with data from other laboratories, our marker set of 17,189 SNPs provides even coverage of the complete worm genome. To date, we have confirmed >1099 evenly spaced SNPs (one every 91 +/- 56 kb) across the six chromosomes and validated the utility of our SNP marker set and new fluorescence polarization-based genotyping methods for systematic and high-throughput identification of genes in C. elegans by cloning several proprietary genes. We illustrate our approach by recombination mapping and confirmation of the mutation in the cloned gene, dpy-18.