The Banni buffaloes distributed in the Kachchh region of Gujarat state in western India are medium to large in body size with a compact body and typical coiled horns. Mean body length, heart girth and height at withers estimated during the survey were 153.7±0.4 cm, 205.5±0.6 cm and 136.7±0.2 cm, respectively. Data on 397 adult milking buffaloes from Banni area based on interviews with farmers, revealed the mean age at first calving to be 39.7±0.4 months, a mean service period of 66.4±1.3 days, a mean lactation length of 293.3±1.5 days and a mean peak yield of 15.7±0.1 litres. Genetic diversity analysis of Banni buffaloes using a set of 15 heterologus bovine microsatellite markers revealed a high degree of allelic polymorphism with a total of 81 alleles and a mean of 5.4. The observed heterozygosity was found to be moderate with a mean of 0.506 and ranged from 0.125 (ILSTS 045) to 0.795 (ILSTS 008). The mean F value (0.181) across 15 microsatellite loci was significantly positive indicating heterozygote deficiency in the population. The test for Hardy-Weinberg equilibrium revealed seven of the 15 microsatellite loci analyzed showing significant deviations.