The study aimed to characterise the genetic diversity of Kangayam, Umblachery, Alambadi, Bargur, Pulikulam, Deoni and Ongole cattle breeds present in South India. A total of 96 animals were genotyped for 27 FAO-recommended microsatellite loci. The genotypes were analysed using various bioinformatics tools. A total of 1340 alleles were observed across the 27 microsatellite markers and 7 breeds, with allele count ranging from 6 to 13. The average number of observed alleles obtained in Alambadi, Bargur, Deoni, Kangayam, Ongole, Pulikulam and Umbalacherry cattle were 6.59 ±2.15, 6.96 ± 2.05, 7.52 ±2.42, 6.19 ±2.32, 6.67 ± 2.11, 8.07 ± 2.13 and 7.63 ± 2.34, respectively. The highest number (Na=14) of alleles was observed in Deoni, whereas the lowest number (Na=3) of alleles was noticed in Ongole, Kangayam and Alambadi breeds of cattle. Most of the microsatellite markers were considered informative (PIC > 0.50) and the PIC values showed a significant positive correlation with the number of alleles (r = .517, p = .006), regardless of differences in allele size. The dendrogram among the seven breeds revealed that the Alambadi and Bargur breeds of cattle formed one cluster, while the Kangayam, Umblacherry and Ongole cattle were distinctly different from all other breeds of cattle.