Previous studies indicated that Asiatic lions and tigers were thought to be highly inbred with very limited genetic diversity. However, our research on these species has revealed significantly greater levels of genetic variation than previously reported. For instance, our analysis using Randomly Amplified Polymorphic DNA (RAPD) on 38 Asiatic lions from the Gir Forest Sanctuary in India showed an average heterozygosity of 25.82% across four primers. This finding was supported by complementary sperm motility studies conducted by our colleagues. In Indian tigers, our examination using microsatellite analysis on 22 individuals and multilocus fingerprinting with the Bkm 2(8) probe demonstrated a heterozygosity of 22.65%. Additionally, microsatellite analysis on hybrids between Asiatic and African lions using loci Fca 77 and Fca 126 showed polymorphism, which has been useful in distinguishing pure Asiatic lions from hybrids. Similar techniques were employed to identify hybrids of Indian and Siberian tigers via PCR amplification of hair samples. To understand the genetic variation before the population bottleneck around the turn of the present century, we conducted microsatellite analysis on skin samples from museum specimens dating back 50 to 125 years. Remarkably, the genetic variability observed in these historical samples (21.01%) was comparable to that found in the present population. This suggests that the low genetic diversity observed in these species may be inherent rather than solely the result of extensive inbreeding. Our DNA fingerprinting studies on Asiatic lions and tigers have been instrumental in identifying individuals with significant genetic diversity, which is crucial for effective conservation breeding programs.