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Comparison of methods to extract PCR-amplifiable DNA from fruit, herbal and black teas

Author

Listed:
  • Eliška Čermáková

    (Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Prague, Czech Republic)

  • Kamila Zdeňková

    (Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Prague, Czech Republic)

  • Kateřina Demnerová

    (Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, University of Chemistry and Technology Prague, Prague, Czech Republic)

  • Jaroslava Ovesná

    (Crop Research Institute, Prague, Czech Republic)

Abstract

The success of polymerase chain reaction (PCR) assay depends on template deoxyribonucleic acid (DNA) being sufficient with respect to both quantity and quality. Some biological materials contain compounds which inhibit the functioning of DNA polymerase and thus need to be removed as part of the DNA extraction procedure. The aim of the present experiments was to optimise the process of DNA isolation from various types of black, fruit and herbal teas. A comparison was made between two cetyltrimethylammonium bromide (CTAB)-based protocols and two commercially available DNA purification kits. The yield and integrity of the extracted DNA were monitored both spectrophotometrically and using agarose gel electrophoresis. The presence/absence of inhibitors in the DNA preparations was checked by running quantitative real-time PCRs. The optimal protocol was deemed to be the CTAB method described in ISO 21571:2005, so this method is recommended for the routine sample analysis of tea products.

Suggested Citation

  • Eliška Čermáková & Kamila Zdeňková & Kateřina Demnerová & Jaroslava Ovesná, 2021. "Comparison of methods to extract PCR-amplifiable DNA from fruit, herbal and black teas," Czech Journal of Food Sciences, Czech Academy of Agricultural Sciences, vol. 39(5), pages 410-417.
    • Handle: RePEc:caa:jnlcjf:v:39:y:2021:i:5:id:24-2021-cjfs
    DOI: 10.17221/24/2021-CJFS
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