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. 2021 Sep;58(9):3561-3567.
doi: 10.1007/s13197-021-05079-4. Epub 2021 Mar 24.

Evaluation of DNA extraction methods for molecular traceability in cold pressed, solvent extracted and refined groundnut oils

Keotshepile Precious Bojang  1 , Aparna Kuna  2 , Sreerangam N C V L Pushpavalli  3 , Supta Sarkar  4 , M Sreedhar  5
Affiliations

Evaluation of DNA extraction methods for molecular traceability in cold pressed, solvent extracted and refined groundnut oils

Keotshepile Precious Bojang et al. J Food Sci Technol. 2021 Sep.

Abstract

Groundnut oil (GNO)/peanut oil is one of the agro-food products with great economic value and hence an attractive target for adulteration and mislabeling. Simple Sequence Repeats (SSR) are markers of choice for DNA fingerprinting studies as they exhibit high polymorphism due to variable number of repeats. Hence, this study was designed to evaluate and optimize a method for DNA isolation from groundnut oil and study the possibility of using the isolated DNA for molecular traceability using SSR markers. Four methods to isolate DNA from groundnut oil were evaluated. All the four methods were modified CTAB protocols, but differed in procedures for extraction, buffer compositions, amount of oil used and DNA carriers. For molecular traceability of oils, extraction and recovery of DNA from edible oil is a key step, especially in refined oils. A method that employed DNA enrichment prior to extraction with CTAB buffer yielded amplifiable DNA from cold pressed GNO, crude hexane extracted GNO and refined GNO. The optimized method for isolation of DNA from groundnut oil is simple, efficient, less costly and reproducible when compared to chromatography and spectroscopy based techniques.

Keywords: DNA extraction; Fingerprinting; Food adulteration; Groundnut oil; SSR markers.

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Conflict of interest statement

Conflict of interestThe authors report no conflicts of interest. The authors alone are responsible for the content, writing of the paper and complete responsibility for the integrity and accuracy of the data presented.

Figures

Fig. 1
Fig. 1
Agarose gel electrophoresis of genomic DNA visualized with ethidium bromide (M4). M-1 kb ladder; Lane 1: positive control (groundnut leaf); Lane 2: CPGNO; Lane 3: HCGNO and Lanes 4–5: CRGNO
Fig. 2
Fig. 2
Agarose gel electrophoresis of PCR products using DNA extracted from GNO oils using the primer GM1573. Lane 1: (100 bp ladder); Lane 2: positive control (Groundnut leaf DNA); Lane 3: CRGNO; Lane 4: HCGNO and Lanes 5–6: CPGNO
Fig. 3
Fig. 3
Agarose gel image of PCR products obtained using DNA isolated from CRGNO with different SSR primers. Lane 1: 100 bp ladder); Lane 2: positive control (groundnut leaf); Lane 3: Primer GM1573; Lane 4: GM1009; Lane 5: GM2301; Lane 6: PM137 and Lane 7: Seq5Do5
Fig. 4
Fig. 4
Agarose gel of PCR products (DNA from CPGNO) using 4 SSR primers. Lane 1: (50 bp ladder); Lane 2: GM1009; Lane 3: GM2301; Lane 4: PM137 and Lane 5: PM137
See this image and copyright information in PMC

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