Research on genetic polymorphism of growth hormone (GH) and receptor growth hormone (rGH) has not been done in crossbred of Limousin cattle, so it is interesting to be examined. Blood samples were taken from 14 Madura calves were artificially inseminated with Limousin cement. DNA amplification is done by using Polymerase Chain Reaction (PCR) method, Restriction Fragment Length Polymorphism (RFLP) method to determine the genotype. DNA sequencing was done to determine nucleotide sequences of GH unit genes. The results showed that identification of GH and rGH gene polymorphisms was done by breaking DNA fragments from 432 and 298 bp in Madura and Limousin cattle (Madrasin) ie, L and V alleles have a frequency of 0.67 and 0.33 for the GH gene, respectively. This proves that the crossed-breeding of Madrasin have V allele that is not owned by the Madura cattle. While in the rGH gene, the A allele is 0.92 and the G allele is 0.08, with the frequency of the A allele larger than the G allele. This research concluded: that GH and rGH undergo changes on polymorphisms in Madrasin cattle can be used as a basis for selection.