title: Absolute quantification of olive oil DNA by droplet digital-PCR (ddPCR): Comparison of isolation and amplification methodologies.
abstract: 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.
authors.affiliation: Instituto de Agricultura Sostenible (IAS-CSIC), Consejo Superior de Investigaciones Científicas, Alameda del Obispo s/n, 14080 Córdoba, Spain; Dipartimento di Agricoltura, Alimentazione e Ambiente (Di3A), Via Valdisavoia 5, Università degli Studi di Catania, 95123 Catania, Italy. Electronic address: fscollo@unict.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Scollo
authors.affiliation: Instituto de Agricultura Sostenible (IAS-CSIC), Consejo Superior de Investigaciones Científicas, Alameda del Obispo s/n, 14080 Córdoba, Spain; Dep. Bioquímica y Biología Molecular, Campus Rabanales C6-1-E17, Campus de Excelencia Internacional Agroalimentario (ceiA3), Universidad de Córdoba, 14071 Córdoba, Spain. Electronic address: z32ayegl@uco.es.
authors.forename: Leticia A
authors.initials: LA
authors.lastname: Egea
authors.affiliation: Dipartimento di Agricoltura, Alimentazione e Ambiente (Di3A), Via Valdisavoia 5, Università degli Studi di Catania, 95123 Catania, Italy; Horticulture and Landscape College, Hunan Agricultural University, Changsha, Hunan, China. Electronic address: gentilea@unict.it.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Gentile
authors.affiliation: Dipartimento di Agricoltura, Alimentazione e Ambiente (Di3A), Via Valdisavoia 5, Università degli Studi di Catania, 95123 Catania, Italy. Electronic address: slamalfa@unict.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: La Malfa
authors.affiliation: Dep. Bioquímica y Biología Molecular, Campus Rabanales C6-1-E17, Campus de Excelencia Internacional Agroalimentario (ceiA3), Universidad de Córdoba, 14071 Córdoba, Spain. Electronic address: bb1dopeg@uco.es.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Dorado
authors.affiliation: Instituto de Agricultura Sostenible (IAS-CSIC), Consejo Superior de Investigaciones Científicas, Alameda del Obispo s/n, 14080 Córdoba, Spain. Electronic address: phernandez@ias.csic.es.
authors.forename: Pilar
authors.initials: P
authors.lastname: Hernandez
mesh_terms: D004247:DNA / Q000235:genetics*; D006801:Humans; D000069463:Olive Oil / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D000069463:Olive Oil; D004247:DNA
keywords: Molecular markers; Olea europaea; Olive oil; Traceability; qRT-PCR