Rapid identification of thousands of copperhead snake (Agkistrodon contortrix) microsatellite loci from modest amounts of 454 shotgun genome sequence

摘要

Optimal integration of next-generation sequencing into mainstream research requires reevaluation of how problems can be reasonably overcome and what questions can be asked. One potential application is the rapid acquisition of genomic information to identify microsatellite loci for evolutionary, population genetic and chromosome linkage mapping research on non-model and not previously sequenced organisms. Here, we report on results using high-throughput sequencing to obtain a large number of microsatellite loci from the venomous snake Agkistrodon contortrix, the copperhead. We used the 454 Genome Sequencer FLX next-generation sequencing platform to randomly sample approximately 26.8 Mbp (128,773 reads) of the copperhead genome, thus sampling about 2% of the genome of this species. We identified microsatellite loci in 11.3% of all reads obtained, with 14,612 microsatellite loci identified in total, 4,564 of which had flanking sequences suitable for PCR primer design. The random sequencing-based approach to identify microsatellites was rapid, cost-effective, and identified thousands of useful microsatellite loci in a previously unstudied species.