Journal
APIDOLOGIE
Volume 44, Issue 2, Pages 163-172Publisher
SPRINGER FRANCE
DOI: 10.1007/s13592-012-0168-3
Keywords
SSRs; cloning; next-generation sequencing; Colletidae
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Funding
- Andrew W. Mellon Foundation at Cornell University
- Sarah Bradley Fellowship
- Cornell Biology Research Fellowship Program
- National Science Foundation [DEB-0814544, DEB-0742998]
- Division Of Environmental Biology
- Direct For Biological Sciences [0814544] Funding Source: National Science Foundation
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The recent implementation of next-generation sequencing for the discovery of microsatellite markers has made this technology the most effective method for generating genetic markers in non-model organisms. Here, we report the de novo discovery of microsatellite markers for the solitary bee Colletes inaequalis using cloning/Sanger sequencing and direct 454 pyrosequencing from microsatellite-enriched genomic libraries. We identified and successfully multiplexed 18 highly variable microsatellite markers in 585 individuals. The number of alleles per locus ranged from 3 to 23, and the expected heterozygosity ranged from 0.056 to 0.912. These genetic markers will allow for the investigation of levels of inbreeding and fine-scale population structure in C. inaequalis. Our results contribute to the literature demonstrating that 454 sequencing is more time- and cost-efficient than cloning/Sanger sequencing at identifying a large number of genomic regions with microsatellite repeat motifs.
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