Abstract: Evaluation of genetic diversity was made using RAPD as a genetic marker system in allohexaploid wheat (T. aestivum). A set of 30 RAPD markers were used for 43 wheat cultivars to examine their utility for detecting DNA polymorphism in genotypes and estimating genetic diversity among them. A total of 228 different RAPD bands were scored for the presence of bands among all the varieties. Out of 30 oligonucleotide primers used, 5 primers gave reproducible DNA product (bands) suitable for the establishment of genetic diversity among the varieties. The Genetic Similarity (GS) coefficient for all pairs of genotypes was ranged from 0.66 to 1.0. V 35 (HD 2747) and V 39 (HI 8498) had high coefficient of similarity (0.982) while, V 4 (VL 804) and V 6 (VL 809) showed less coefficient of similarity (0.646). It indicated that V 35 (HD 2747) and V 39 (HI 8498) carry closely related ancestry while the cultivars V 4 (VL 804) and V 6 (VL 809) are not close to each other. Present results revealed the potential of RAPD technique in determining genetic diversity and make suitable fingerprints for wheat cultivars.