The burying beetle Nicrophorus americanus Olivier is an endangered species known to occur in disjunct populations in 6 states. Parsimony and maximum likelihood analysis of the nuclear ribosomal DNA first internal transcribed spacer (ITS1) sequences from 10 Nicrophorinae species revealed N. americanus to form a distinct clade with N. orbicollis Say. Genetic variation within and among 5 N. americanus populations, collected from South Dakota, Nebraska, Oklahoma, Arkansas, and Rhode Island, was studied. Ribosomal DNA ITS1 sequences from 14 beetles revealed 48 polymorphic and 20 informative nucleotide sites. N. americanus genetic divergence was between 0.16 and 4.76%. We found little evidence that these 5 populations have maintained unique genetic variation. No nucleotide sites were found that were diagnostic for any of the 5 populations examined, indicating that these populations may not be necessarily treated as separate, independent objects of conservation. However, further genetic investigation is warranted before translocations are attempted among the remaining populations of the American burying beetle.