Absolute quantification of olive oil DNA by droplet digital-PCR (ddPCR): Comparison of isolation and amplification methodologies

Authors：Scollo Francesco^*; Egea Leticia A; Gentile Alessandra; La Malfa Stefano; Dorado Gabriel; Hernandez Pilar

Source：Food Chemistry, 2016, 213: 388-394.

DOI：10.1016/j.foodchem.2016.06.086

• Olea europaea
• Olive oil
• Traceability
• Molecular markers
• qRT-PCR

Olive oil is considered a premium product for its nutritional value and health benefits, and the ability to define its origin and varietal composition is a key step towards ensuring the traceability of the product. However, isolating the DNA from such a matrix is a difficult task. In this study, the quality and quantity of olive oil DNA, isolated using four different DNA isolation protocols, was evaluated using the qRT-PCR and ddPCR techniques. The results indicate that CTAB-based extraction methods were the best for unfiltered oil, while Nucleo Spin-based extraction protocols showed greater overall reproducibility. The use of both qRT-PCR and ddPCR led to the absolute quantification of the DNA copy number. The results clearly demonstrate the importance of the choice of DNA-isolation protocol, which should take into consideration the qualitative aspects of DNA and the evaluation of the amplified DNA copy number.

• Institution
  6; 1; 湖南农业大学; 5