Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH2O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×103 copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields.
| Published in | Journal of Plant Sciences (Volume 10, Issue 3) |
| DOI | 10.11648/j.jps.20221003.11 |
| Page(s) | 86-90 |
| 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 |
Rf1 Event, Loop-Mediated Isothermal Amplification, Genetically Modified Organism, Sensitivity, Specificity
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APA Style
Zhang Jihong, Wang Jiaying, Zhao Lei, Huang Suwen, Chen Xianfeng. (2022). Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). Journal of Plant Sciences, 10(3), 86-90. https://doi.org/10.11648/j.jps.20221003.11
ACS Style
Zhang Jihong; Wang Jiaying; Zhao Lei; Huang Suwen; Chen Xianfeng. Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). J. Plant Sci. 2022, 10(3), 86-90. doi: 10.11648/j.jps.20221003.11
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Download@article{10.11648/j.jps.20221003.11,
author = {Zhang Jihong and Wang Jiaying and Zhao Lei and Huang Suwen and Chen Xianfeng},
title = {Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.)},
journal = {Journal of Plant Sciences},
volume = {10},
number = {3},
pages = {86-90},
doi = {10.11648/j.jps.20221003.11},
url = {https://doi.org/10.11648/j.jps.20221003.11},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20221003.11},
abstract = {Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH2O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×103 copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields.},
year = {2022}
}
TY - JOUR T1 - Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.) AU - Zhang Jihong AU - Wang Jiaying AU - Zhao Lei AU - Huang Suwen AU - Chen Xianfeng Y1 - 2022/05/07 PY - 2022 N1 - https://doi.org/10.11648/j.jps.20221003.11 DO - 10.11648/j.jps.20221003.11 T2 - Journal of Plant Sciences JF - Journal of Plant Sciences JO - Journal of Plant Sciences SP - 86 EP - 90 PB - Science Publishing Group SN - 2331-0731 UR - https://doi.org/10.11648/j.jps.20221003.11 AB - Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH2O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×103 copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields. VL - 10 IS - 3 ER -
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