This research was carried out from June to November 2017. This research aims to analyze genetic relationship of tilapia (Oreochromis niloticus) nirwana strain which is cultured in Aceh Besar (Aceh Province) and in Wanayasa (West Java Province) with RAPD-PCR technique. The DNA Isolation Activity of tilapia nirwana strain was carried out in The Biotechnology Laboratory, Building No.4, FPIK UNPAD, Indonesia and the PCR process was carried out in The Microbiology and Biotechnology Laboratory, Building No.3 FPIK UNPAD, Indonesia. The purpose of this research is to determine the level of genetic relationship of tilapia nirwana strain cultured in Aceh Besar and Wanayasa and to find out whether tilapia nirwana strain in Aceh Besar can be used as a genetic stock. The results showed that genetic relationship level of tilapia nirwana strain cultured in Aceh Besar with those cultured in Wanayasa is 67%. The existence of environmental differences results in genetic diversity between Aceh Besar and Wanayasa tilapia nirwana strain. Factors that causing grouping movement are external factors such as: Geographical isolation and habitat fragmentation as well as internal factors such as mutation, natural selection, genetic drift, and gene flow. Tilapia nirwana strain that is cultured in Aceh Besar produce polymorphic bands, so it can be used as a new genetic stock to stabilize the tilapia nirwana strain production.