Genetic polymorphisms of μ-calpain, a calcium-dependent protease specifically expressed in muscle and related to meat tenderness, have been investigated in cattle worldwide, but not well documented in Indonesian beef cattle. This study was done to evaluate the frequency of SNP 316 in the μ-calpain gene in Aceh cattle, a local Indonesian beef cattle plays important role in the fulfillment of red meat for peoples in Aceh and its neighboring areas. For this purpose, genomic DNA was isolated from 29 sirloin meats samples collected from cull, female aceh cattle slaughtered at the Slaughter House of Banda Aceh. The cattle were purposively selected based on physical characteristics and hair colors referred to Indonesian law. For the SNP genotyping, PCR-RFLP methods were set up. The results showed all three genotypes namely CC, CG, and GG were found in Aceh cattle. The allele and genotype frequencies of SNP CAPN316 in the CAPN gene were: C – 0.14 and G – 0.86; CC – 3.4%, CG – 20.7% and GG – 75.9%. The observed and expected frequencies of CAPN SNP 316 in the cattle population examined were GG 22.0 and 21.6, CG 6.0 and 6.9, and CC 1.0 and 0.6.