A Cervus genotyping kit based on automated fluorescent multiplex PCR for rapid characterization of genetic diversity in several deer populations
2000
Bonnet, A. | Thevenon, Sophie | Maudet, Frédéric | Maillard, Jean-Charles
Microsatellite loci are highly conserved among related species (Kühn et al, 1996) such as cattle and sheep (Moore et al, 1992). Transfer of a large number of polymorphic microsatellites between families of the Artiodactyla order has been demonstrated several times (Engel et al, 1996; Kuhn et al, 1996, Slate et al, 1998 and Roed, 1998). Therefore, a multi-allele system should be useful in studying the genetic diversities of populations. This study was developed using four different deer species (Rusa, Eld, Swamp and Vietnamese Sika deer). A set of 38 microsatellites derived from bovine and ovine origin were chosen based on two criteria: (i) known to amplify in other deer species (Red deer); and (ii) showing an interesting polymorphic level as described in previous studies (Slate et al, 1998 and Talbot et al, 1996). From these 38 screened markers, 30 gave an amplified product in the 4 deer species (78.9%). Of those 30 markers, 14-20 (40%-60%) were polymorphic, depending on the species. Using 12 microsatellites polymorphic in the 4 species, we set up a unique multiplex PCR optimized for annealing temperature and reagent concentrations. The 12 primer sets were labelled with 3 different fluorochromes, depending on the allelic range for each species. The automatic analysis was performed using an ABI 377 sequencer and PE Genotyper software. This method for parentage testing or genotyping gave good and reproducible results for the 4 studied species as well as for several other tested deer species and subspecies. This tool could be considered a first generation "Cervus" genotyping kit useful for the rapid characterization of genetic diversity. (Résumé d'auteur)
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