SPECIES IDENTIFICATION INFERRED THROUGH MITOCHONDRIAL 16S rRNA GENE SEQUENCE BETWEEN FRESHWATER TESTUDINES - LISSEMYS PUNCTATA AND MELANOCHELYS TRIJUGA.

Lalitha, R.; Chandavar., VidyaR

doi:10.21474/ijar01/4964

Cited by 2 publications

(1 citation statement)

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“…16S rRNA holds high inter-species DNA variations and low intra-species DNA variation (Taniguchi et al, 2022), providing high confidence in meat species discrimination. Within the mitochondrial genes, 16S rRNA has been used for broad range of mammalian and birds' species because of its evolutionary stability (Vences et al, 2016;Ha et al, 2017;Lalitha & Chandavar, 2017). Furthermore, the developed direct PCR method is coupled with RFLP assay which is much more desirable for molecular based meat identification, especially, in cases where large number of samples have to be processed as it does not require DNA sequencing and/or specialized equipment and reduces the cost and time for post-PCR processing of samples (Guan et al, 2019;Gargouri et al, 2021;Vaithiyanathan et al, 2021;Taha et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

Comparison of different DNA preparatory methods for development of a universal direct PCR-RFLP workflow for identification of meat origin in food products

et al. 2023

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The quality and quantity of the extracted DNA are two key aspects for a successful PCR (Polymerase Chain Reaction) amplification. Moreover, reduction in time and cost required for DNA extraction are also two considerable factors, in cases when large number of samples are to be analyzed within a limited time-frame and budget. Accordingly, the aim of this study was to compare and optimize performance of five different DNA extraction methods by boiling meat tissues from cattle, buffalo, sheep, goat, chicken, camel, horse and dog in PBS (Phosphate Buffer Saline), distilled water, alkaline lysis buffers 1, 2 or 3. The results indicated that the boiling of meat and its products in alkaline lysis buffers was a good method to extract crude DNA. The optimized crude DNA extraction protocol was coupled with PCR-RFLP (Restriction Fragment Length Polymorphism) analysis for meat species differentiation. This developed workflow was tested on fifty-three commercial beef and mutton samples, out of which three samples were found to be adulterated. In conclusion, the rapid crude DNA extraction protocol has led to the development of a direct PCR-RFLP workflow that is simple, time-saving and cost-effective for PCR-based identification of different meat species.

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