Quantification of DNA fragmentation in processed foods using real-time PCR
| Autor: | Shin-ichi Fukudome, Hiroyuki Kawakami, Reona Takabatake, Kazumi Kitta, Reiko Teshima, Akio Noguchi, Takuya Hayashida, Youichi Kurimoto, Junichi Mano, Kazunari Kondo, Yasuyuki Nishitsuji, Yosuke Kikuchi |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
DNA
Plant Base pair Food Genetically Modified Computational biology DNA Fragmentation Biology Real-Time Polymerase Chain Reaction 01 natural sciences Polymerase Chain Reaction Analytical Chemistry chemistry.chemical_compound 0404 agricultural biotechnology Fragmentation (cell biology) Gene business.industry 010401 analytical chemistry 04 agricultural and veterinary sciences General Medicine Ribosomal RNA 040401 food science 0104 chemical sciences Genetically modified organism Biotechnology Real-time polymerase chain reaction chemistry DNA fragmentation business DNA Food Science |
| Zdroj: | Food chemistry. 226 |
| ISSN: | 1873-7072 |
| Popis: | DNA analysis of processed foods is performed widely to detect various targets, such as genetically modified organisms (GMOs). Food processing often causes DNA fragmentation, which consequently affects the results of PCR analysis. In order to assess the effects of DNA fragmentation on the reliability of PCR analysis, we investigated a novel methodology to quantify the degree of DNA fragmentation. We designed four real-time PCR assays that amplified 18S ribosomal RNA gene sequences common to various plants at lengths of approximately 100, 200, 400, and 800 base pairs (bp). Then, we created an indicator value, "DNA fragmentation index (DFI)", which is calculated from the Cq values derived from the real-time PCR assays. Finally, we demonstrated the efficacy of this method for the quality control of GMO detection in processed foods by evaluating the relationship between the DFI and the limit of detection. |
| Databáze: | OpenAIRE |
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