Isolation and characterization of microsatellite loci for use in population genetic analysis in the timber rattlesnake, Crotalus horridus.

A Crotalus horridus genomic library was screened for clones containing microsatellite loci by hybridization with oligonucleotides consisting of a (dC x dA)n dinucleotide repeat. Primers designed to amplify six of the microsatellite loci were used to screen 32 unrelated individuals representing populations in eastern Pennsylvania, southern New Jersey, North Carolina, South Carolina, and Alabama. The six microsatellite loci were all polymorphic, with two to nine alleles, and heterozygote frequencies at each locus from 0.1 to 0.69. Allelic frequencies varied among geographically separated populations. Screening of two families produced no evidence of multiple paternity. These microsatellite markers should be useful for the assessment of kinship relationships and genetic diversity within and between populations of C. horridus. The application of this technology could provide a valuable tool for the development of effective conservation and management programs for threatened and endangered populations.