Background: PPARGC1A gene mediates expression of genes involved in gluconeogenesis and is regarded as a prominent candidate gene governing the effect of milk fat in dairy bovines. The study was planned to identify DNA polymorphisms in the PPARGC1A gene in Gaolao Cattle and to find their association with milk production traits in Gaolao cattle. Methods: The blood was collected from 224 purebred Gaolao cattle from farm and field along with test day milk from each cow for estimating milk components. Genomic DNA was assessed using phenol-chloroform extraction method and Kits. PCR-RFLP, PCR-SSCP and direct DNA sequencing methods were used to identify polymorphism. Result: PPARGC1AG1- NheI locus of 3ʹUTR region were polymorphic with allele frequency for A alleles as 0.84 and for allele B as 0.16. In studied animals, significant differences were not observed for milk traits between genotypes at PPARGC1A-NheI locus. The analysis at PPARGC1AG4-SSCP revealed no significant differences in milk traits; however sequencing revealed SNP. PPARGC1AG1- NheI genotype-based sequencing revealed T greater than C SNP at 263rd position 02 computational SNPs viz; C-T at 263rd position and A-T at 268th position. The identified variation at the PPARGC1A gene may aid in identifying associations for development of markers for selection and genetic improvement.