Using next-generation DNA sequencing for rapid microsatellite discovery in Mexican leaf-toed geckos (Phyllodactylus tuberculosus)

作者：Blair Christopher^*; Jimenez Arcos Victor H; Mendez de la Cruz Fausto R; Murphy Robert W

来源：Conservation Genetics Resources, 2012, 4(3): 807-810.

DOI：10.1007/s12686-012-9649-y

• Evolution
• Gecko
• Genomics
• Pyrosequencing

We use a combination of 454 shotgun pyrosequencing and traditional molecular cloning to isolate microsatellite loci for the Mexican yellow-bellied leaf-toed gecko (Phyllodactulus tuberculosus). A pyrosequencing run on a 454 GS Junior yields 62 Mbp of data composed of 150 k fragments with an average length of 412 bp. Among the fragments, 18 144 (12 %) contain a suitable microsatellite. Dinucleotides and tetranucleotides are most frequently encountered. We then genotype 30 individuals of P. tuberculosus using 12 polymorphic loci. We find moderate allelic diversity and heterozygosity. No loci are in linkage disequilibrium and only two loci show deviations from Hardy-Weinberg expectations. Our results further highlight the utility of next-generation sequencing for discovering molecular markers in non-model taxa and suggest that sequencing on a 454 GS Junior system is a rapid and cost-effective approach for microsatellite characterization.

• 出版日期2012-9