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Tytuł:
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Simple, Precise, and Less Biased GMO Quantification by Multiplexed Genetic Element-Specific Digital PCR.
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Autorzy:
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Noma S; Research Center for Basic Science, Nisshin Seifun Group Inc, 5-3-1 Tsurugaoka, Fujimino, Saitama 356-8511, Japan.
Kikuchi Y; Research Center for Basic Science, Nisshin Seifun Group Inc, 5-3-1 Tsurugaoka, Fujimino, Saitama 356-8511, Japan.
Satou M; Food Research Center, Nippn Corporation, 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Tanaka T; Food Research Center, Nippn Corporation, 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Takiya T; Food Research Center, Nippn Corporation, 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Okusu H; Food Research Center, Nippn Corporation, 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Futo S; FASMAC Co., Ltd, 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Takabatake R; Institute of Food Research, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Kitta K; Institute of Food Research, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Mano J; Institute of Food Research, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
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Źródło:
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Journal of AOAC International [J AOAC Int] 2022 Feb 04; Vol. 105 (1), pp. 159-166.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Publication: 2020- : Oxford : Oxford University Press
Original Publication: Arlington, VA : AOAC International, c1992-
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MeSH Terms:
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Multiplex Polymerase Chain Reaction*
Zea mays*/genetics
DNA ; DNA, Plant/genetics ; Plants, Genetically Modified/genetics ; Real-Time Polymerase Chain Reaction
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Substance Nomenclature:
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0 (DNA, Plant)
9007-49-2 (DNA)
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Entry Date(s):
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Date Created: 20211009 Date Completed: 20220210 Latest Revision: 20220210
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Update Code:
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20240104
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DOI:
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10.1093/jaoacint/qsab138
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PMID:
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34626115
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Background: To provide the consumer with choices of genetically modified organisms (GMO) or non-GMO, official food labeling systems were established in many countries. Because the threshold GMO content values were set to distinguish between "non-GMO" and "GMO" designations, GMO content quantification methods are required for ensuring the appropriateness of labeling.
Objective: As the number of GMOs is continuously increasing around the world, we set out to develop a low-cost, simple and less biased analytical strategy to cover all necessary detection targets.
Methods: Digital PCR methods are advantageous compared to the conventional quantitative real-time PCR methods. We developed a digital PCR-based GMO quantification method to evaluate the GMO content in maize grains. To minimize the analytical workload, we adopted multiplex digital PCR targeting the 35S promoter and the nopaline synthase terminator, which are genetic elements commonly introduced in many GMOs.
Results: Our method is significantly simpler and more precise than the conventional real-time PCR-based methods. Additionally, we found that this method enables quantification of the copy number of GMO DNA without double counting multiple elements (35S promoter and nopaline synthase terminator) tandemly placed in a recombinant DNA construct.
Conclusion: This is the first report on the development of a genetically modified maize quantification method using a multiplexed genetic element-specific digital PCR method. The tandem effect we report here is quite useful for reducing the bias in the analytical results.
Highlights: Multiplexed genetic element-specific digital PCR can simplify weight-based GMO quantification and thus should prove useful in light of the continuous increase in the number of GM events.
(© AOAC INTERNATIONAL 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
