The horse is renowned for its racing performance, gaitedness, dancing and other personality traits. On the bases of these aesthetic characteristics, this valued species of Equus caballus is an integral part of different sports. A number of studies showing its racing performance is associated with MSTN gene in different horse populations of the world. So, the current study aimed to explore the genetic variability of 18:66493737T>C variant in the Pakistani random-bred horse population too. ARMS-PCR was used to genotype a total of 24 horses taken from equine clinics. Our initial results showed that overall, 54% population is homozygous wild type (TT), 41% is heterozygous (TC) and the remaining 4% is homozygous mutant (CC) at this locus. Alternative allele frequency within the best performers and control horses was 0.36 & 0.12 respectively. Similarly, Chi-Square association test was applied using PLINK data toolset which gave us a significant p-value of 7.832 × 10-6. Hence, the Pakistani horse population also proposes the variability of this locus in conformity to other horse populations of the world. Furthermore, similar studies may also be intended with racing performance data to evaluate different indigenous horse breeds with a large sample size, and subject marker may be propagated to other breeds by adopting targeted breeding strategies.