This study utilized micro-morphological identification and microsatellite SSLP DNA genotyping to characterize subterranean termites collected from 73 locations across the state of Nebraska. Micro-morphological characters of soldier termites from 30/73 collection sites revealed the presenceof two species, Reticulitermes flavipes (Kollar) and R. tibialis Banks. PCR based microsatellite genotyping was undertaken at nine loci for all sample sites. PCR products were resolved by 2.5% agarose gel electrophoresis, stained with ethidium bromide, and visualized by UV illuminesence. Comparison of micro-morphological species assignment to the genotyping data for workers from the 30 soldier sample sites revealed that PCR-based genotyping reliably seperated the samples into genotypic groups that corresponded with morphological characterization. Genotypic analysis of workers from all 73 sample sites revealed four informative loci which enabled the grouping of sites into R. flavipes, R. tibialis, and a mixed genotypic group. Data from both characterizations were used to construct termite distribution maps to enable possible future research to focus on the zones of cohabitation between the two species.