Abstract: RAPD markers were used to fingerprint thirty hexaploid wheat varieties (Triticum aestivum L.) from different genetic background. A total of 76 DNA fragments were amplified, using five primers, ranging in size from 1884 to 200 base pairs. Out of the 76 amplified products, 19.7% were monomorphic and 80.3% were polymorphic, averaging 12.2 polymorphisms per primer. The number of DNA fragments for each primer varied from 9 (OP-B14) to 20 (OP-C15) with an average of 15.2 fragments per primer. One specific marker out of 11 amplified fragments was detected for primer OP-B11 and two specific markers out of 9 amplified fargments were detected for primer OP-B14. The genetic similarity between varieties ranged from 32 to 97%, with an average of 64.5%. The large genetic diversity may be due to the different areas from which the genotypes were collected. Some distinctive varieties showed high genetic similarity with other varieties, such as Sids1 and Sids 8 (97%), Sids 7 and Icarda 46 (96%), Icarda1 and Gimeza 9 (95% ). On the contrary, some varieties displayed low genetic similarity such as Giza164 and Sakha 60 (32%). The similarity values showed clearly substantial differences among the wheat varieties. The dendrogram resulting from the UPGMA cluster analysis showed that the studied varieties could be divided into two main clusters from the same node. The first cluster contained ten varieties four of them are Egyptian, while the second cluster contained 19 varieties including 13 Egyptian. The dendrogram clustered the genotypes into ten groups and showed efficiency in identifying genetic variability. The results indicated that RAPD analysis is useful as molecular genetic marker for estimating the genetic diversity between wheat varieties