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Page No. : 71-74

ABSTRACT

The aim of this study was to develop species-specific polymerase chain reaction (PCR) assay for species detection of beef using self-designed primer pair based on D-loop region of mitochondrial gene. The annealing temperature for PCR amplification of designed primer pair was optimized and primer pair was tested for its specificity using DNA template from cattle, buffalo, sheep, goat and pig. The primer pair was also subjected to sensitivity test by serial dilution of DNA template and specificity was validated in known and mixed meat samples. The developed PCR assay successfully amplified the DNA template to a fragment length of 399bp specific to cattle at optimized annealing temperature of 62°C. The primer pair was able to amplify minimum of 10 ng DNA template and successfully identified the genesis of tissue in known and meat admixed samples. It was concluded that the developed PCR assay was highly species specific, sensitive and useful for detection of tissue of cattle origin.

Keywords: cattle, D-loop, PCR, specificity, sensitivity