Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol.13, Issue 4, 2011; Page No.(643-646)

DETECTION OF SPECIES SPECIFIC MILK BY USING POLYMERASE CHAIN REACTION

ANUPAM SINGH, ASHWANI KUMAR SINGH, JITENDER SINGH AND RAGHVENDAR SINGH

Abstract

The adulteration of food products is a significant problem in the food processing and production. Most frequently, such products are adulterated that are produced in big quantities and further, the expensive products whose adulteration brings a profit. This is how fraudulent producers try to cheat consumers and authorities. Adulteration of costly milk with cheaper one becomes a matter of concern for research workers. Buffalo milk is frequently used for adulteration because of its prevailing production in the world and its lower price as compared to other types. The present study was carried out for detection of milk species with the use of cytochrome b gene variability by PCR. Milk samples from Buffalo, sheep and goat were utilized for molecular analysis. The milks of these principal dairying species are well characterized. Genomic DNA was isolated from four samples of each species. Mitochondrial cytochrome b gene was amplified by PCR using a common forward primer and speciesspecific reverse primer. PCR cycling protocol included initial denaturation at 94deg.C for 5 minutes then followed by 40 cycles of 94deg.C for 30 seconds, 60deg.C for 30 seconds 72deg.C for 30 seconds and final extension at 72deg.C for 10 minutes. PCR amplicons were resolved by Agarose gel electrophoresis and for each species produce a characteristic band pattern. The PCR products showed species-specific DNA fragments of 155 bp, 270 bp, 330 bp from goat, Buffalo and sheep respectively and will help traders and consumers against milk adulteration.

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