Comparative study of DNA extraction methods from fresh and processed yellowfin tuna muscle tissue

Piskatá, Zora; Pospisilova, Eliska; Bořilová, Gabriela

doi:10.1080/10942912.2017.1297953

Cited by 28 publications

(28 citation statements)

References 27 publications

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“…The DNA was considered to be satisfactorily pure when the ratios of the A260 to A280 were within the range of 1.7-2.0. DNA contamination with proteins usually reduces the A260 to A280 ratio to values lower than 1.7 (Cawthorn et al 2011;Piskata et al 2017). DNA is very sensitive to acid and alkaline agents, because of the mechanism of hydrolytic degradation of DNA (Peano et al 2004;Chapela et al 2007), and low pH media have been described as favouring a higher DNA degradation (Bauer et al 2003;Chapela et al 2007).…”

Section: Discussionmentioning

confidence: 99%

A quantitative comparison of two kits for DNA extraction from canned tuna

et al. 2019

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The most common methods that can be used for species identification of tuna include methods based on detection of species-specific DNA via the polymerase chain reaction (PCR) method. The problem with DNA detection in processed products is the possibility of DNA fragmentation during the technological process. The quantity and quality of extracted DNA is a crucial step for species identification based on the DNA analysis. In this study, two DNA extraction methods (DNeasy Blood & Tissue Kit and DNeasy mericon Food Kit) for tuna DNA isolation were compared. Eight food products of canned tuna (three of them were declared as Thunnus albacares and five products were declared as Katsuwonus pelamis) with a different addition of various ingredients were tested. Furthermore, three different times of proteolysis (30 min, 60 min, overnight) for each sample and each extraction kit were evaluated. The DNA concentration was determined by a Qubit dsDNA HS Assay Kit fluorescence method and quantified using a Qubit fluorometer. The DNA purity was evaluated using the A260/A280 ratio of absorbances measured on a spectrophotometer. The main indicator of DNA quality and quantity was its amplifiability in the subsequent real-time PCR for Thunnus species, Thunnus albacares and Katsuwonus pelamis. Based on the results, both kits can be used for tuna species determination in highly heat-treated products with different composition, nevertheless, the DNeasy mericon Food Kit provided better statistical values in some parameters. The effect of different times of proteolysis was significant in most of the samples with regard to the crossing point values determined by real-time PCR.

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Section: Discussionmentioning

confidence: 99%

A quantitative comparison of two kits for DNA extraction from canned tuna

et al. 2019

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“…Moreover, DNA is largely unaffected by tissue source or sample damage [4,5]. However, fragment size is a limiting factor for the subsequent PCR [6]. Thus, DNA-based analytical methods represent a crucial approach for species identification in raw and highly processed meat products due to their high thermal stability and their very low detection limits.…”

Section: Introductionmentioning

confidence: 99%

“…which considerably influence the quality of DNA [19,20,21,22], it is necessary to individually optimize DNA isolation procedures for each type of food product. In addition, the chemical compounds present in food matrices (polysaccharides, proteins, collagen, polyphenols, fulvic acids, or lipids) may not be completely removed during the DNA extraction protocol and can affect the integrity of DNA or cause inhibition of subsequent PCR analysis [6]. Inhibitor compounds can interfere with PCR by decreasing or even completely inhibiting the activity of DNA polymerase [23].…”

Section: Introductionmentioning

confidence: 99%

The Quality of DNA Isolated from Processed Food and Feed via Different Extraction Procedures

et al. 2019

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The extraction of DNA is a critical step for species identification by PCR analysis in processed food and feed products. In this study, eight DNA extraction procedures were compared—DNeasy Blood and Tissue Kit, DNeasy mericon Food Kit, chemagic DNA Tissue 10 Kit, Food DNA Isolation Kit, UltraPrep Genomic DNA Food Mini Prep Kit, High Pure PCR Template Preparation Kit, phenol—chloroform extraction, and NucleoSpin Food—Using self-prepared samples from both raw and heat-processed and/or mechanically treated muscles and different types of meat products and pet food (pork, beef, and chicken). The yield, purity, and suitability of DNA for PCR amplification was evaluated. Additionally, comparisons between the effectiveness of various extraction methods were made with regard to price, and labor- and time-intensiveness. It was found that the DNeasy mericon Food Kit was the optimal choice for the extraction of DNA from raw muscle, heat-treated muscle, and homemade meat products from multiple and single species.

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“…DNA concentration is also affected by the ingredients of the mixtures contained in the product. The additions of polysaccharides, proteins, or lipids and other ingredients in processed products cause the isolated DNA mix with contaminant compounds (Piskata et al 2017). Maulid and Nurilmala (2015) reported that the processed mackerel cracker DNA product could not be observed on agarose gel electrophoresis due to low concentration.…”

Section: Dna Concentration and Puritymentioning

confidence: 99%

Advancing traceability using DNA sequence on seafood product: fraudulence in shrimp crackers

Nurilmala¹,

Ajitama²,

Jacoeb³

et al. 2020

Biodiversitas

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Abstract. Nurilmala M, Atjitama Y, Jacob AM, Indarwati AR, Nugraha R. 2020. Advancing traceability using DNA sequence on seafood product: fraudulence in shrimp crackers. Biodiversitas 21: 5650-5656. Shrimp crackers are snacks made from starch with the addition of shrimp and other ingredients. Authentication of shrimp crackers becomes a quality control method in line with its traceability since the morphological characters are not recognized in those products. The failure of the authentication will lead to adulteration of shrimp products. The study aimed to design shrimp-specific polymerase chain reaction (PCR) primers by and used them to authenticate shrimp cracker products to assure their traceability. The primers were designed from cytochrome oxidase subunit I (COI) gene with a GC content of 50% and a melting temperature of 57 °C. The amplicon had a length of 613 bp. Fourteen shrimp crackers having DNA concentrations 0.3 to 8.1 ng/µL were evaluated. Eight out of fourteen DNA samples were successfully amplified using the specific primer and could be visualized on the agarose gel at 500-750 bp. Those DNA samples were identified belonging to shrimp species, namely Litopenaeus vannamei, Metapenaeus ensis, and Fenneropenaeus merguiensis with 95% to 100% identity. Meanwhile, the other six DNA samples underwent PCR using universal primers. One sample was amplified and identified as fish (Sphyraena flavicauda). These results indicate illegal substitution of shrimp using unidentified species occurred in shrimp crackers.

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Comparative study of DNA extraction methods from fresh and processed yellowfin tuna muscle tissue

Cited by 28 publications

References 27 publications

A quantitative comparison of two kits for DNA extraction from canned tuna

A quantitative comparison of two kits for DNA extraction from canned tuna

The Quality of DNA Isolated from Processed Food and Feed via Different Extraction Procedures

Advancing traceability using DNA sequence on seafood product: fraudulence in shrimp crackers

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