title: Next-generation sequencing as a tool to quickly identify causative EMS-generated mutations.
abstract: The advent of next generation sequencing has influenced every aspect of biological research. Many labs are now using whole genome sequencing in Arabidopsis thaliana as a means to quickly identify EMS-generated mutations present in isolated mutants. Following identification of these mutations, examination of T-DNA insertional alleles defective in candidate genes or complementation of the mutant phenotype with a wild type copy of candidate genes can be used to verify which mutation is causative for the phenotype of interest. Here, we discuss the benefits and pitfalls of using this method to identify mutations underlying phenotypes.
authors.affiliation: a Department of Biology; St. Louis University , St. Louis , MO , USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Thole
authors.forename: L C
authors.initials: LC
authors.lastname: Strader
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D005020:Ethyl Methanesulfonate; D016296:Mutagenesis
chemical_list: D005020:Ethyl Methanesulfonate
keywords: ABA, abscisic acid; AR, ABA Root Resistance; EMS, ethylmethane sulfonate; NGS, next-generation sequencing; PCR, polymerase chain reaction; SNP, single nucleotide polymorphism; abscisic acid; ethylmethane sulfonate; mutation mapping; next-generation sequencing