Abstract
We use a combination of 454 shotgun pyrose- quencing 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 microsat- ellite. Dinucleotides and tetranucleotides are most fre- quently encountered. We then genotype 30 individuals of P. tuberculosus using 12 polymorphic loci. We find mod- erate 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.
Published Version
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