DNA isolation was done by the 8 different cyst populations collected from the potato growing areas of Aegean Region (Turkey) between 2006-2008. As a result of Multiplex PCR conducted with species-specific primer, composing of band is observed in 435 bp as specific to Globodera rostochiensis Wollenweber (Tylenchida: Heteroderidae). By doing PCR with rDNA2 and rDNA1.58S primers of the same sample, as a consequence of cropping with Hinf I enzyme, in 233bp and 522bp band were confirmed as specific to G. rostochiensis. In this study, other potato cyst nematodes, Globodera pallida Stone, 1973 was not determined. Globodera rostochiensis was determined previously by using morphometric methods in Turkey. The identification of G. rostochiensis was confirmed by molecular methods with this study.