Development of nucleic acid isolation by non-silica-based nanoparticles and real-time PCR kit for edible vegetable oil traceability

Abstract

For efficient extraction of amplifiable DNA from edible vegetable oils, we developed a novel DNA extraction approach based on the non-silica-based dipolar nanocomposites. The nanoparticle comprises a hydrophilic polymethyl methacrylate core with abundant capillaries, hydrophilic vesicles decorated with molecules having DNA affinity and a coating hydrophobic polystyrene layer. The nanoparticles are soluble in oil, adsorb the DNA from the aqueous phase and gave a high DNA recovery ratio. All DNA extracts from fully refined vegetable oil soybean, peanut, rapeseed, and cottonseed oils, including their blends, were sufficiently pure to be amplified by real-time PCR targeting the chloroplast ribulose-1,5-bisphosphate gene (rbcL), therefore, the species of origin and their ratios in mixed vegetable oils blended from two or three oil-species could be determined. These results indicate that the novel DNA isolation and real-time PCR kit is a simple, sensitive and efficient tool for the species identification and traceability in refined vegetable oils.

Keywords: 2-Mercaptoethanol (PubChem CID: 1567); Chloroform (PubChem CID: 6212); DNA extraction; Edible vegetable oils; Ethylenediaminetetraacetic acid (PubChem CID: 6049); Guanidine thiocyanate (PubChem CID: 135742599); Isopropanol (PubChem CID: 3776); Methyl methacrylate (PubChem CID: 6658); Non-silicon based dipolar nanocomposites; Polyvinyl pyrrolidone (PubChem CID: 131751496); Real-time PCR; Sodium dodecyl sulfate (PubChem CID: 3423265); Styrene (PubChem CID: 7501); Traceability; Triton X-100 (PubChem CID: 5590).