Lentil (Lens culinaris Medik.subsp. culinaris) is an important principal cool season pulse crop of Nepal. To estimate genetic diversity, one hundred eighty five diverse lentil germplasm were collected from National Grain Legume Research Program, Rampur; Regional Agricultural Research Station, Nepalgunj and National Agriculture Genetic Resource Center, Khumaltar. Thirty polymorphic microsatellites marker were used for PCR analysis and finger printing. The results obtained from microsatellite profiling revealed that maximum alleles were amplified from SSR 19, SSR 99, SSR 113, SSR 156 and SSR 202 with amplicon size 180-395. Highly informative and detectable polymorphic markers for this study, foundwere SSR 34-2, SSR 90 and SSR 207 which indicate the power and higher resolution of those marker systems in detecting molecular diversity. The dendrogram constructed by highly polymorphic 30 SSR markers from 185 lentil accessions showed ten major groups from Group I to X based on source of origin and their pedigree. Groups III and VI genotypes were totally different from other groups whereas group X genotypes were from Nepal cross lines. This study show the divergency among lentil genotypes which can be further used in lentil breeding programs. All genotypes involved in this study exhibited wide range of genetic variability due to different center of origin, different genetic constitution. The level of genetic relatedness largely depends on the type of molecular markers used in the study, nature of SSR repeat motif, number of SSR markers and the genetic relatedness of the lentil germplasm to be analysed. The genetic relatedness detected in this study may constitute the foundation for future systematic lentil breeding programs.