Lentil (Lens culinaris ssp. culinaris) is consumed in many countries as a rich source of protein in largely vegetarian diets. Australia grows lentil as a cash crop in rotation with cereal and produces predominantly red lentils that are exported throughout the world, particularly to countries in South Asia and the Middle East. Differentiation of varieties is important when exporting products to such markets, maintaining variety purity during seed production and in the collection of end-point royalties. Lentil-specific and fluorescent sequenced tagged microsatellite markers (STMS) markers were used to construct a DNA fingerprint database for 10 Lens culinaris ssp. culinaris genotypes (Northfield, Digger, ILL7537, Nugget, Indianhead, ILL2024, ILL6788, Palouse, Nipper and Boomer) that represent major new cultivars and key breeding lines within the Australian breeding program. All 10 lentil genotypes were distinguished using the assessed STMS loci. Unique alleles were observed for several lines, including Boomer and Nipper, varieties recently released in Australia. This database will play an important role in seed typing for commercial export certification and the commercial management of cultivars.