Abstract
The PCR-RFLP using 16S rRNA gene was explored to test its effectiveness in identification of speciation of meat from porcine in raw, cooked as well as admixed meat. Critical perusal of the restriction maps of ∼591-bp 16S rRNA fragment sequences from pig as well as other livestocks identified RsaI specific to pig, yielding 343-bp and 251-bp fragments in pig, while in other livestock species such as cattle, buffalo, goat and sheep, no restriction enzyme site was found. In all the samples of raw pork, the restriction digestion of 594-bp fragment with RsaI yielded the 343-bp and 251-bp fragments, while no restriction digestion in all other livestock species, i.e. cattle, buffalo, goat and sheep. PCR-RFLP with RsaI (In cursive) of 594-bp fragment amplified from three different types of cooked meat DNA samples from pig, i.e. cooking at 72°C for 30 min, steam cooking at 90°C for 30 min and autoclaving at 120°C/15 lb/30 min, yielded a similar restriction enzyme digestion profile as with raw meat DNA, suggesting no effect of cooking on PCR-RFLP. Similarly, PCR-RFLP was able to differentiate the origin of meat in mixed meat samples quite consistently up to the level of 25%; however, at the level below 10%, results were not consistent.
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Sharma, A., Kumar, S., Sharma, N., Sharma, D. (2012). PCR-RFLP of 16S rRNA Gene for Identification of Speciation of Meat of Porcine Origin in Processed and Admixed Meat. In: Sabu, A., Augustine, A. (eds) Prospects in Bioscience: Addressing the Issues. Springer, India. https://doi.org/10.1007/978-81-322-0810-5_6
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DOI: https://doi.org/10.1007/978-81-322-0810-5_6
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