SRA

We are developing a library of 2,000 mutagenized strains and sequencing each to a depth of 15x as a community resource. From previous estimates this should yield a library of half to one million SNP’s with 5-10 non-synonymous changes per average gene across the collection. After testing various mutagens we are using EMS, ENU and a cocktail of EMS plus ENU. In the F1 we select for unc-22 mutations to ensure that the genome has been mutagenized. In the F2 we select for non-unc-22 animals and then these animals are self-crossed for 10 generations to ensure that the final isolate is homozygous across all regions of the genome. We will supplement these strains with natural isolates to recover additional mutations. For sequencing we load size-selected, barcoded samples on either a GAII, or Hi-Seq sequencing machine (Illumina) and do paired-end reads. Data analysis employs phaster, SamTools and custom filters. Sequence data will be deposited in WormBase and the individual strains will be available from the Caenorhabditis Genetics Center for detailed study. Eventually we hope to distribute the 2,000 strains as a single kit, allowing parallel experimentation on a wide spectrum of mutant genes.