title: Rapid mapping and identification of mutations in Caenorhabditis elegans by restriction site-associated DNA mapping and genomic interval pull-down sequencing.
abstract: Forward genetic screens provide a powerful approach for inferring gene function on the basis of the phenotypes associated with mutated genes. However, determining the causal mutation by traditional mapping and candidate gene sequencing is often the rate-limiting step, especially when analyzing many mutants. We report two genomic approaches for more rapidly determining the identity of the affected genes in Caenorhabditis elegans mutants. First, we report our use of restriction site-associated DNA (RAD) polymorphism markers for rapidly mapping mutations after chemical mutagenesis and mutant isolation. Second, we describe our use of genomic interval pull-down sequencing (GIPS) to selectively capture and sequence megabase-sized portions of a mutant genome. Together, these two methods provide a rapid and cost-effective approach for positional cloning of C. elegans mutant loci, and are also applicable to other genetic model systems.
authors: Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
authors: Sean M
authors: SM
authors: O'Rourke
authors: John
authors: J
authors: Yochem
authors: Amy A
authors: AA
authors: Connolly
authors: Meredith H
authors: MH
authors: Price
authors: Luke
authors: L
authors: Carter
authors: Joshua B
authors: JB
authors: Lowry
authors: Douglas W
authors: DW
authors: Turnbull
authors: Nick
authors: N
authors: Kamps-Hughes
authors: Nicholas
authors: N
authors: Stiffler
authors: Michael R
authors: MR
authors: Miller
authors: Eric A
authors: EA
authors: Johnson
authors: Bruce
authors: B
authors: Bowerman
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D056426:Genetic Loci / Q000235:genetics; D016678:Genome / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics; D015183:Restriction Mapping / Q000191:economics / Q000379:methods*
chemical_list: D004247:DNA