Most adulteration is a problem in the meat industry entailing economic, ethical and human health concerns.  This study aimed to develop a PCR assay that can detect meat adulteration through the use of species-specific primers.  Testing of species-specific primers by Matsunaga et al. (1999) showed cross amplification to non-target species, hence, a need to design new primers with higher specificity.  To design species-specific primers that can discriminate cattle from carabao and horse meat, the Cytochrome B gene of cattle was cloned and sequenced.  Cloning the CytB was done through Rapid Alkaline Lysis DNA extraction, PCR amplification using Naidu et al. (2012) primers, ligation with pGem-T Easy cloning vector, and cell transformation via heat shock.  Transformed colonies were selected, and the plasmids with insert were purified then sequenced.  The generated consensus sequence of the cattle CytB was aligned with published sequences of horse and carabao CytB; the species-specific regions were located and used for the design of primers.  In silico analysis showed that new primers were more specific than the Matsunaga et al. (1999) primers in terms of the specificity of the both forward and reverse primers, the length of the primers, and the 3’ end mismatching to non-target organisms.  In vitro testing of the primers is still needed to validate its specificity.