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Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms

The rapid increase in the number of genetically modified (GM) varieties has led to a demand for high-throughput methods to detect genetically modified organisms (GMOs). We describe a new dynamic array-based high throughput method to simultaneously detect 48 targets in 48 samples on a Fludigm system....

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Detalles Bibliográficos
Autores principales: Li, Xiaofei, Wu, Yuhua, Li, Jun, Li, Yunjing, Long, Likun, Li, Feiwu, Wu, Gang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5154595/
https://www.ncbi.nlm.nih.gov/pubmed/25556930
http://dx.doi.org/10.1038/srep07616
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author Li, Xiaofei
Wu, Yuhua
Li, Jun
Li, Yunjing
Long, Likun
Li, Feiwu
Wu, Gang
author_facet Li, Xiaofei
Wu, Yuhua
Li, Jun
Li, Yunjing
Long, Likun
Li, Feiwu
Wu, Gang
author_sort Li, Xiaofei
collection PubMed
description The rapid increase in the number of genetically modified (GM) varieties has led to a demand for high-throughput methods to detect genetically modified organisms (GMOs). We describe a new dynamic array-based high throughput method to simultaneously detect 48 targets in 48 samples on a Fludigm system. The test targets included species-specific genes, common screening elements, most of the Chinese-approved GM events, and several unapproved events. The 48 TaqMan assays successfully amplified products from both single-event samples and complex samples with a GMO DNA amount of 0.05 ng, and displayed high specificity. To improve the sensitivity of detection, a preamplification step for 48 pooled targets was added to enrich the amount of template before performing dynamic chip assays. This dynamic chip-based method allowed the synchronous high-throughput detection of multiple targets in multiple samples. Thus, it represents an efficient, qualitative method for GMO multi-detection.
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spelling pubmed-51545952016-12-20 Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms Li, Xiaofei Wu, Yuhua Li, Jun Li, Yunjing Long, Likun Li, Feiwu Wu, Gang Sci Rep Article The rapid increase in the number of genetically modified (GM) varieties has led to a demand for high-throughput methods to detect genetically modified organisms (GMOs). We describe a new dynamic array-based high throughput method to simultaneously detect 48 targets in 48 samples on a Fludigm system. The test targets included species-specific genes, common screening elements, most of the Chinese-approved GM events, and several unapproved events. The 48 TaqMan assays successfully amplified products from both single-event samples and complex samples with a GMO DNA amount of 0.05 ng, and displayed high specificity. To improve the sensitivity of detection, a preamplification step for 48 pooled targets was added to enrich the amount of template before performing dynamic chip assays. This dynamic chip-based method allowed the synchronous high-throughput detection of multiple targets in multiple samples. Thus, it represents an efficient, qualitative method for GMO multi-detection. Nature Publishing Group 2015-01-05 /pmc/articles/PMC5154595/ /pubmed/25556930 http://dx.doi.org/10.1038/srep07616 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Li, Xiaofei
Wu, Yuhua
Li, Jun
Li, Yunjing
Long, Likun
Li, Feiwu
Wu, Gang
Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms
title Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms
title_full Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms
title_fullStr Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms
title_full_unstemmed Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms
title_short Development and Validation of A 48-Target Analytical Method for High-throughput Monitoring of Genetically Modified Organisms
title_sort development and validation of a 48-target analytical method for high-throughput monitoring of genetically modified organisms
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5154595/
https://www.ncbi.nlm.nih.gov/pubmed/25556930
http://dx.doi.org/10.1038/srep07616
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