Detection threshold of the cytochrome B gene in meat samples

Wong, Annie Fui Ling (2005) Detection threshold of the cytochrome B gene in meat samples. Universiti Malaysia Sabah. (Unpublished)


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The morphological ambiguities regarding to the traditional methods of animal species classification has lead to the requirement of a more reliable and practical method of species identification. A polymerase chain reaction-based analysis has been developed by amplification of the conserved region of the mitochondrial DNA (mtDNA) cytochrome b (cyt b) gene using universal primers CYTB 1 and CYTB2. A good quality of total DNA was extracted from mixtures of raw chicken and turkey and also beef and buffalo and was subsequently subjected to PCR amplification yielding distinct bands of approximately 360 bp in all species. The threshold detection of the mixtures was determined by RFLP analysis by selected restriction enzymes (RsaI, Bstnl, TaqI and AluI). RsaI has successfully discriminated both chicken and turkey resulting a detection sensitivity limit of 5% turkey. While beef and buffalo was differentiated by two restriction enzymes Bstnl and TaqI producing fragments of 318bp and 42bp for buffalo and 191 bp and 168bp for beef. AluI has failed to produce variable RFLP pattern in this mixture. However, the sensitivity of the beef and buffalo mixture with beef as the minor component cannot be determined as species substitution might have occurred due to the similar banding pattern observed in the gel where beef was suspected to be mislabeled with buffalo. The cyt b PCR-RFLP threshold detection of raw meat samples has proved to be sufficient in both sensitivity and species identification assay.

Item Type: Academic Exercise
Uncontrolled Keywords: meat sample, threshold detection, enzyme, banding pattern, species identification
Subjects: Q Science > QD Chemistry
Divisions: SCHOOL > School of Science and Technology
Date Deposited: 11 Jul 2013 05:29
Last Modified: 23 Oct 2017 07:06

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