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Abstract

This research aims to determine the genetic diversity of Leptin exon 2 and intron 2 in Pesisir cattle using sequencing methods. The samples used in this study consist of 96 blood samples from Pesisir cattle raised at BPTU Padang Mengatas, Payakumbuh, West Sumatra. Blood samples were isolated using the genomic DNA Purification Kit (Promega) protocol. The isolated DNA was then amplified using a pair of forward primers 5’- ATC TGA AGA CCT GGA TGC GG -3’ and reverse primers 5’- TGG TTG CAG GTC TGA GCT TA -3’ resulting in a Leptin gene exon 2 and intron 2 fragment with a length of 744 bp. Then the amplification products were sent for sequencing with the assistance of 1st Base Singapore’s services. The research results indicate that there are 13 diversities in the intron 2 region of the Pesisir cattle samples, with the discovery of 12 mutations occurring at positions +327 G?A, +398 C?T, +399 A?G, +439 C?T, +441 C?A, +547 G?C, +565 C?G, +622 G?A, +629 C?T, +647 C?T, +693 T?C, +712 G?T and 1 deletion at position +751 G?del. In the Hardy-Weinberg equilibrium test, there are 8 mutations that are in Hardy-Weinberg equilibrium, which are at positions +327, +399, +439, +547, +565, +622, +693, +712 while 4 mutations are not in Hardy-Weinberg equilibrium, which are at positions +398, +441, +629 dan +647. In this research, a check of restriction enzymes revealed the presence of 12 restriction enzymes that can be used in studies utilizing the PCR-RFLP (Polymerase Chain Reaction – Restriction Fragment Length Polymorphism) method.