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India is mega diversity center for buffalo breeds among that Mehsana is one of the best milk breeds of buffalo. Diacylglycerol acyltransferase 1 (DGAT1) is a microsomal enzyme that catalyzes the final step of triglyceride synthesis. The DGAT1 gene is a strong functional candidate for determining milk fat content. In this work, we used different techniques viz., Polymerase chain reaction – restriction fragment length polymorphism (PCRRFLP), Allele specific PCR (AS PCR), Single extension assay (SNaP Shot) and direct sequencing to screen the nineteen reported single nucleotide polymorphisms (SNPs) in DGAT1 gene in 130 unrelated Mehsana buffalo (Bubalus bubalis) reared under field progeny testing programme operated by Dudhsagar Research and Development Association (DURDA) in the Gujarat. SNP g.8259G>A was significantly associated with first lactation yield. However, no SNP was associated with milk fat content in Mehsana buffalo. The result presented here is preliminary and require further investigation on large population.