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A Comparative Study of Some Procedures for Isolation of Fruit DNA of Sufficient Quality for PCR-Based Assays

Faculty of Chemistry, Brno University of Technology, 612 00 Brno, Czech Republic
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Academic Editor: Carmen Cuadrado
Molecules 2020, 25(18), 4317; https://doi.org/10.3390/molecules25184317
Received: 11 August 2020 / Revised: 18 September 2020 / Accepted: 18 September 2020 / Published: 20 September 2020
Food fraud has been and still is a problem in the food industry. It is detectable by several approaches, such as high performance liquid chromatography (HPLC), chemometric assays, or DNA-based techniques, each with its own drawbacks. This work addresses one major drawback of DNA-based methods, in particular their sensitivity to inhibitors contained in particular matrices from which DNA is isolated. We tested five commercial kits and one in-house method characterized by different ways of sample homogenization and DNA capture and purification. Using these methods, DNA was isolated from 10 different fruit species commonly used in plant-based foodstuffs. The quality of the DNA was evaluated by UV-VIS spectrophotometry. Two types of qPCR assays were used for DNA quality testing: (i) Method specific for plant ITS2 region, (ii) methods specific for individual fruit species. Based mainly on the results of real-time PCR assays, we were able to find two column-based kits and one magnetic carrier-based kit, which consistently provided fruit DNA isolates of sufficient quality for PCR-based assays useful for routine analysis and identification of individual fruit species in food products. View Full-Text
Keywords: DNA isolation; real-time PCR; commercial kit; red fruit; Prunus; tropical fruit DNA isolation; real-time PCR; commercial kit; red fruit; Prunus; tropical fruit
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MDPI and ACS Style

Fialova, L.; Romanovska, D.; Marova, I. A Comparative Study of Some Procedures for Isolation of Fruit DNA of Sufficient Quality for PCR-Based Assays. Molecules 2020, 25, 4317. https://doi.org/10.3390/molecules25184317

AMA Style

Fialova L, Romanovska D, Marova I. A Comparative Study of Some Procedures for Isolation of Fruit DNA of Sufficient Quality for PCR-Based Assays. Molecules. 2020; 25(18):4317. https://doi.org/10.3390/molecules25184317

Chicago/Turabian Style

Fialova, Lenka, Denisa Romanovska, and Ivana Marova. 2020. "A Comparative Study of Some Procedures for Isolation of Fruit DNA of Sufficient Quality for PCR-Based Assays" Molecules 25, no. 18: 4317. https://doi.org/10.3390/molecules25184317

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