Reintroductions are an important conservation and management technique used to restore extirpated populations. Negative genetic consequences (e.g., diversity loss, bottlenecks, inbreeding) are often an unintentional result of reintroductions, due to a small number of founders or suboptimal habitat at release sites. American martens (Martes americana) were extirpated from Michigan's Lower Peninsula in 1911 due to habitat loss and unregulated trapping. Martens were reintroduced into 2 areas of the Lower Peninsula in 1985–1986. The Lower Peninsula reintroduction was characterized by a relatively small number of founders (85 individuals) released into 2 geographically disparate, fragmented sites. We genotyped martens sampled at the 2 release sites approximately 20–25 years since reintroduction, using 11 microsatellite loci. We detected low average allelic richness (3.92 alleles per locus), moderate levels of inbreeding (mean F_IS = 0.106), and multiple loci with significant heterozygote deficiencies. Effective population size estimates were small, ranging between 6 and 27 individuals depending on the estimator and the sample group. We also detected significant population structuring between the release sites (F_ST = 0.093 using the most recent sample). With small population size and limited to no gene flow, we predict the 2 Lower Peninsula marten populations will continue to diverge and potentially further lose genetic diversity. This study highlights the importance of long-term genetic monitoring of reintroduced populations.