Medicinal plants are an important source of compounds for the pharmaceuticals and other industries and traditional medicine. Tinospora cordifolia, commonly known as giloe, is one of the important medicinal climbers which belongs to the family Menispermaceae. Studies were undertaken for identification and genetic variation within six accessions of T. cordifolia through random amplified polymorphic DNA (RAPD) markers. Analysis was made using 12 primers. Out of them, eight primers showed amplification and were selected for genetic relationship within six accessions. A total of 81 bands were produced of size 100 to 950 bp with an average of 6.75 bands for each primer, of which 80 bands were polymorphic. The polymorphism percentage was 98.56 and Polymorphic Index Content (PIC) value ranged from 0.722 to 0.992. The genetic similarity index was calculated using Jaccard's similarity coefficient which ranged from 0.4225 to 0.7000. The dendrogram was constructed using similarity matrix based on UPGMA method, which resulted into two main clusters. The two genotypes TC2 and TC3 were closely related accessions, whereas TC4 and TC6 were distantly related accessions. In conclusion, clustering of majority of accessions in one group reflected the narrow genetic structure of Tinospora populations in the region and the RAPD technique is highly popular and reveals relatively high levels of polymorphism.