The importance of germplasm characterization is an important link between the conservation and utilization of plant genetic
resources in various breeding programmes. In the present study, genetic variability and relationships among 25 Lactuca
sativa L. genotypes were tested using random amplified polymorphic DNA (RAPD) molecular markers. A total of 45 random
decamer oligonucleotide primers were examined to generate RAPD profiles, out of these reproducible patterns were obtained
with 22 primers. A total of 87 amplicon were obtained, out of which all were polymorphic and 7 were unique bands. The
level of polymorphism across genotypes was 100% as revealed by RAPD. Genetic similarity matrix, based on Jaccard’s
coefficients ranged from 13.7 to 84.10% indicating a wide genetic base. Dendrogram was constructed by unweighted pair
group method with arithmetic averages method. RAPD technology could be useful for identification of different accessions
as well as assessing the genetic similarity among different genotypes of lettuce. The study reveals the limited genetic base
and the needs to diversify using new sources from the germplasm.