Visual Detection of Rf2 Event in Transgenic Rapeseed (<i>Brassica napus</i> L.) Using Loop-Mediated Isothermal Amplification

Wang Jiaying; Zhao Lei; Li Wen; Huang Suwen; Zhang Jihong

doi:doi:10.11648/j.jps.20221006.14

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Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification

Wang Jiaying, Zhao Lei, Li Wen, Huang Suwen, Zhang Jihong

Published in Journal of Plant Sciences (Volume 10, Issue 6)

Received: 19 October 2022 Accepted: 2 November 2022 Published: 29 November 2022

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Abstract

Specific methods have been developed in order to efficiently detect GMOs (genetically modified organisms) in both feed and food. Current approaches often rely on thermal cycling devices such as PCR amplifier, which makes it difficult for applications in the wild. Therefore a visual LAMP (Loop-Mediated Isothermal Amplification) method for rapid screening Rf2 event in transgenic rapeseed (Brassica napus L.) is established in this paper. For primer design, position 167-366 in the right border junction sequence of B. napus trangenic line Rf2 (accession number: EU090197.1) was chosen as the amplicon. Reaction mixture (at the volume of 25 μL) consisted of 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 5.0 μL ddH₂O. Dye (SYBR Green I) was pre-added onto the tube lid. The reaction tube was incubated at 60°C for 60 min, followed by heating at 80°C for 10 min to end it. To mix dye with reaction mixture, the tube was centrifuged for 60 s at 8,000 r/min. Corresponding analysis results indicate that this LAMP assay is highly specific and sensitive (1.15×103 copies/μL). In one word, the visual LAMP method specific for Rf2 event in rapeseed, which turns out to be simple, time-effective, sensitive without relying on expensive instruments, is suitable for quick screening in ports.

Published in	Journal of Plant Sciences (Volume 10, Issue 6)
DOI	10.11648/j.jps.20221006.14
Page(s)	222-226
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2022. Published by Science Publishing Group

Keywords

Brassica Napus, Rf2 Event, LAMP, Quick Screening, Ports

References

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APA Style

Wang Jiaying, Zhao Lei, Li Wen, Huang Suwen, Zhang Jihong. (2022). Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification. Journal of Plant Sciences, 10(6), 222-226. https://doi.org/10.11648/j.jps.20221006.14

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ACS Style

Wang Jiaying; Zhao Lei; Li Wen; Huang Suwen; Zhang Jihong. Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification. J. Plant Sci. 2022, 10(6), 222-226. doi: 10.11648/j.jps.20221006.14

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AMA Style

Wang Jiaying, Zhao Lei, Li Wen, Huang Suwen, Zhang Jihong. Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification. J Plant Sci. 2022;10(6):222-226. doi: 10.11648/j.jps.20221006.14

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@article{10.11648/j.jps.20221006.14,
  author = {Wang Jiaying and Zhao Lei and Li Wen and Huang Suwen and Zhang Jihong},
  title = {Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification},
  journal = {Journal of Plant Sciences},
  volume = {10},
  number = {6},
  pages = {222-226},
  doi = {10.11648/j.jps.20221006.14},
  url = {https://doi.org/10.11648/j.jps.20221006.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20221006.14},
  abstract = {Specific methods have been developed in order to efficiently detect GMOs (genetically modified organisms) in both feed and food. Current approaches often rely on thermal cycling devices such as PCR amplifier, which makes it difficult for applications in the wild. Therefore a visual LAMP (Loop-Mediated Isothermal Amplification) method for rapid screening Rf2 event in transgenic rapeseed (Brassica napus L.) is established in this paper. For primer design, position 167-366 in the right border junction sequence of B. napus trangenic line Rf2 (accession number: EU090197.1) was chosen as the amplicon. Reaction mixture (at the volume of 25 μL) consisted of 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 5.0 μL ddH2O. Dye (SYBR Green I) was pre-added onto the tube lid. The reaction tube was incubated at 60°C for 60 min, followed by heating at 80°C for 10 min to end it. To mix dye with reaction mixture, the tube was centrifuged for 60 s at 8,000 r/min. Corresponding analysis results indicate that this LAMP assay is highly specific and sensitive (1.15×103 copies/μL). In one word, the visual LAMP method specific for Rf2 event in rapeseed, which turns out to be simple, time-effective, sensitive without relying on expensive instruments, is suitable for quick screening in ports.},
 year = {2022}
}

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TY - JOUR
T1 - Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification
AU - Wang Jiaying
AU - Zhao Lei
AU - Li Wen
AU - Huang Suwen
AU - Zhang Jihong
Y1 - 2022/11/29
PY - 2022
N1 - https://doi.org/10.11648/j.jps.20221006.14
DO - 10.11648/j.jps.20221006.14
T2 - Journal of Plant Sciences
JF - Journal of Plant Sciences
JO - Journal of Plant Sciences
SP - 222
EP - 226
PB - Science Publishing Group
SN - 2331-0731
UR - https://doi.org/10.11648/j.jps.20221006.14
AB - Specific methods have been developed in order to efficiently detect GMOs (genetically modified organisms) in both feed and food. Current approaches often rely on thermal cycling devices such as PCR amplifier, which makes it difficult for applications in the wild. Therefore a visual LAMP (Loop-Mediated Isothermal Amplification) method for rapid screening Rf2 event in transgenic rapeseed (Brassica napus L.) is established in this paper. For primer design, position 167-366 in the right border junction sequence of B. napus trangenic line Rf2 (accession number: EU090197.1) was chosen as the amplicon. Reaction mixture (at the volume of 25 μL) consisted of 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 5.0 μL ddH2O. Dye (SYBR Green I) was pre-added onto the tube lid. The reaction tube was incubated at 60°C for 60 min, followed by heating at 80°C for 10 min to end it. To mix dye with reaction mixture, the tube was centrifuged for 60 s at 8,000 r/min. Corresponding analysis results indicate that this LAMP assay is highly specific and sensitive (1.15×103 copies/μL). In one word, the visual LAMP method specific for Rf2 event in rapeseed, which turns out to be simple, time-effective, sensitive without relying on expensive instruments, is suitable for quick screening in ports.
VL - 10
IS - 6
ER -

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Author Information

Wang Jiaying

Technical Center, Ningbo Customs, Ningbo, China
Zhao Lei

Ningbo Institute of Inspection and Quarantine Science and Technology, Ningbo, China
Li Wen

Department of Horticultural Technology, Ningbo City College of Vocational Technology, Ningbo, China
Huang Suwen

Technical Center, Ningbo Customs, Ningbo, China
Zhang Jihong

Technical Center, Ningbo Customs, Ningbo, China

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Plain Text BibTeX RIS

APA Style

Wang Jiaying, Zhao Lei, Li Wen, Huang Suwen, Zhang Jihong. (2022). Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification. Journal of Plant Sciences, 10(6), 222-226. https://doi.org/10.11648/j.jps.20221006.14

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ACS Style

Wang Jiaying; Zhao Lei; Li Wen; Huang Suwen; Zhang Jihong. Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification. J. Plant Sci. 2022, 10(6), 222-226. doi: 10.11648/j.jps.20221006.14

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AMA Style

Wang Jiaying, Zhao Lei, Li Wen, Huang Suwen, Zhang Jihong. Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification. J Plant Sci. 2022;10(6):222-226. doi: 10.11648/j.jps.20221006.14

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@article{10.11648/j.jps.20221006.14,
  author = {Wang Jiaying and Zhao Lei and Li Wen and Huang Suwen and Zhang Jihong},
  title = {Visual Detection of Rf2 Event in Transgenic Rapeseed (Brassica napus L.) Using Loop-Mediated Isothermal Amplification},
  journal = {Journal of Plant Sciences},
  volume = {10},
  number = {6},
  pages = {222-226},
  doi = {10.11648/j.jps.20221006.14},
  url = {https://doi.org/10.11648/j.jps.20221006.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20221006.14},
  abstract = {Specific methods have been developed in order to efficiently detect GMOs (genetically modified organisms) in both feed and food. Current approaches often rely on thermal cycling devices such as PCR amplifier, which makes it difficult for applications in the wild. Therefore a visual LAMP (Loop-Mediated Isothermal Amplification) method for rapid screening Rf2 event in transgenic rapeseed (Brassica napus L.) is established in this paper. For primer design, position 167-366 in the right border junction sequence of B. napus trangenic line Rf2 (accession number: EU090197.1) was chosen as the amplicon. Reaction mixture (at the volume of 25 μL) consisted of 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 5.0 μL ddH2O. Dye (SYBR Green I) was pre-added onto the tube lid. The reaction tube was incubated at 60°C for 60 min, followed by heating at 80°C for 10 min to end it. To mix dye with reaction mixture, the tube was centrifuged for 60 s at 8,000 r/min. Corresponding analysis results indicate that this LAMP assay is highly specific and sensitive (1.15×103 copies/μL). In one word, the visual LAMP method specific for Rf2 event in rapeseed, which turns out to be simple, time-effective, sensitive without relying on expensive instruments, is suitable for quick screening in ports.},
 year = {2022}
}

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