title: SNP discovery by mismatch-targeting of Mu transposition.
abstract: Single nucleotide polymorphisms (SNPs) represent a valuable resource for the mapping of human disease genes and induced mutations in model organisms. SNPs may become the markers of choice also for population ecology and evolutionary studies, but their isolation for non-model organisms with unsequenced genomes is often difficult. Here, we describe a rapid and cost-effective strategy to isolate SNPs that exploits the property of the bacteriophage Mu transposition machinery to target mismatched DNA sites and thereby to effectively detect polymorphic loci. To demonstrate the methodology, we isolated 164 SNPs from the unsequenced genome of the Glanville fritillary butterfly (Melitaea cinxia), a much-studied species in population biology, and we validated 24 of them. The strategy involves standard molecular biology techniques as well as undemanding MuA transposase-catalyzed in vitro transposition reactions, and it is applicable to any organism.
authors.affiliation: Metapopulation Research Group, Department of Biological and Environmental Sciences, University of Helsinki, Finland.
authors.forename: Luisa
authors.initials: L
authors.lastname: Orsini
authors.forename: Maria
authors.initials: M
authors.lastname: Pajunen
authors.forename: Ilkka
authors.initials: I
authors.lastname: Hanski
authors.forename: Harri
authors.initials: H
authors.lastname: Savilahti
mesh_terms: D000818:Animals; D020137:Base Pair Mismatch; D002080:Butterflies / Q000235:genetics; D023281:Genomics; D005838:Genotype; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods*; D019895:Transposases / Q000378:metabolism*
chemical_list: C114697:mu transposase; D019895:Transposases