title: High-throughput microsatellite isolation through 454 GS-FLX Titanium pyrosequencing of enriched DNA libraries.
abstract: Microsatellites (or SSRs: simple sequence repeats) are among the most frequently used DNA markers in many areas of research. The use of microsatellite markers is limited by the difficulties involved in their de novo isolation from species for which no genomic resources are available. We describe here a high-throughput method for isolating microsatellite markers based on coupling multiplex microsatellite enrichment and next-generation sequencing on 454 GS-FLX Titanium platforms. The procedure was calibrated on a model species (Apis mellifera) and validated on 13 other species from various taxonomic groups (animals, plants and fungi), including taxa for which severe difficulties were previously encountered using traditional methods. We obtained from 11,497 to 34,483 sequences depending on the species and the number of detected microsatellite loci ranged from 199 to 5791. We thus demonstrated that this procedure can be readily and successfully applied to a large variety of taxonomic groups, at much lower cost than would have been possible with traditional protocols. This method is expected to speed up the acquisition of high-quality genetic markers for nonmodel organisms.
authors.affiliation: INRA, UMR 1301 IBSV INRA/UNSA/CNRS, 400 Route des Chappes, BP 167, 06903 Sophia-Antipolis Cedex, France. thibaut.malausa@sophia.inra.fr
authors.forename: Thibaut
authors.initials: T
authors.lastname: Malausa
authors.forename: André
authors.initials: A
authors.lastname: Gilles
authors.forename: Emese
authors.initials: E
authors.lastname: Meglécz
authors.forename: Hélène
authors.initials: H
authors.lastname: Blanquart
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Duthoy
authors.forename: Caroline
authors.initials: C
authors.lastname: Costedoat
authors.forename: Vincent
authors.initials: V
authors.lastname: Dubut
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pech
authors.forename: Philippe
authors.initials: P
authors.lastname: Castagnone-Sereno
authors.forename: Christophe
authors.initials: C
authors.lastname: Délye
authors.forename: Nicolas
authors.initials: N
authors.lastname: Feau
authors.forename: Pascal
authors.initials: P
authors.lastname: Frey
authors.forename: Philippe
authors.initials: P
authors.lastname: Gauthier
authors.forename: Thomas
authors.initials: T
authors.lastname: Guillemaud
authors.forename: Laurent
authors.initials: L
authors.lastname: Hazard
authors.forename: Valérie
authors.initials: V
authors.lastname: Le Corre
authors.forename: Brigitte
authors.initials: B
authors.lastname: Lung-Escarmant
authors.forename: Pierre-Jean G
authors.initials: PJ
authors.lastname: Malé
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Ferreira
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Martin
mesh_terms: D000818:Animals; D001516:Bees / Q000235:genetics*; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D018895:Microsatellite Repeats; D058889:Molecular Typing / Q000379:methods*
chemical_list: D004247:DNA