Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato

유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색

  • 신원선 (한국식품개발연구원 안전성연구팀) ;
  • 김명희 (한국식품개발연구원 안전성연구팀)
  • Published : 2003.08.01

Abstract

To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

Keywords

GMO labelling;potato;DNA extraction method;PCR detection

References

  1. Meyer, R., Chardonnens. E, Huber, P. and Luthy, J. Polymerase chain reaction (PCR) in the quality and safety assurance of food: detection of soya in processed meat products. Z. Lebnsm. Unters. Forsch. 203: 339-344 (1996) https://doi.org/10.1007/BF01231072
  2. Hubner, P., Studer, E. and Luthy, J. Quantitative competitive PCR for the detection of genetically modified organisms in food. Food Control 10: 353-358 (1999) https://doi.org/10.1016/S0956-7135(99)00074-2
  3. Kim, D.H. Polysaccharide in food, pp. 255-267. In: Food Chemistry, Tamgudang Inc., Seoul, Korea (1992)
  4. Ministry of Agriculture and Forest. MAF's GM-food Labelling Standards. Ministry of Agriculture and Forest. No. 2000-31, Seoul, Korea (2000)
  5. KFDA. Biotech Labelling Standards for Processed Foods (revised). Korea Food and Drug Administration, No. 2001-43, Seoul, Korea (200 I)
  6. Wurz, A., Bluth, A., Zeitz, P., Pfeifer, C. and Willmund, D. Quantitative analysis of genetically modified organisms (GMO) in processed food by PCR-based methods. Food Control 10: 385-389 (1999) https://doi.org/10.1016/S0956-7135(99)00080-8
  7. Brodmann, P. DNA-extraction: Important step of the quantitative detection of GMO in food matrices. New Food 5:57-61 (2001)
  8. Gilbert, J. Sampling of raw materials and processed foods for the presence of GMOs. Food Control 10: 363-365 (1999) https://doi.org/10.1016/S0956-7135(99)00076-6
  9. KFDA. Guideline of Detection Method for Genetically Modified Food. Korea Food and Drug Administration, Seoul, Korea (2002)
  10. Vaitilingom, M., Pijinenburg, H., Gendre, F. and Brignon, P. Realtime quantitative PCR detection of genetically modified maximizer maize and roundup ready soybean in some representative foods. J. Agric. Food Chem. 47: 5261-5266 (1999) https://doi.org/10.1021/jf981208v
  11. Kaufman, B., Richards, S. and Dierig, D. DNA isolation method for high polysaccharide Lesqurella species. Industrial Crops Products 9: 111-114 (1999) https://doi.org/10.1016/S0926-6690(98)00021-1
  12. KFDA. Biotech Labelling Standards for Processed Foods. Korea Food and Drug Administration, No. 2000-43, Seoul, Korea (2000)
  13. Jaccaud, E., Hohne, M. and Meyer, R. Assessment of screening methods for the identification of genetically modified potatoes in raw materials and finished products. J. Agric. Food Chem. 51: 550-557 (2003) https://doi.org/10.1021/jf0208031
  14. Vollenhofer, S., Burg, K., Schmidt, J. and Kroath, H. Genetically modified organisms in food-screening and specific detection by polymerase chain reaction. J. Agric. Food Chem. 47: 5038-5043 (1999) https://doi.org/10.1021/jf990353l
  15. Kuiper, H.A. Summary report of the ILSI Europe workshop on detection methods for novel foods derived from genetically moditied organisms. Food Control 10: 339-349 (1999) https://doi.org/10.1016/S0956-7135(99)00072-9
  16. James, C. Global review of commercialized transgenic crops: 2001, ISAAA briefs No. 24. pp. 1-31, Ithaca, NY (2001)