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Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences

BACKGROUND: The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR). Here we have applied the loop-mediated isothermal amplification (LAMP) method to amplify GMO-related DNA sequences, 'internal' common...

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Detalles Bibliográficos
Autores principales: Lee, David, La Mura, Maurizio, Allnutt, Theo R, Powell, Wayne
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2656497/
https://www.ncbi.nlm.nih.gov/pubmed/19187544
http://dx.doi.org/10.1186/1472-6750-9-7
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author Lee, David
La Mura, Maurizio
Allnutt, Theo R
Powell, Wayne
author_facet Lee, David
La Mura, Maurizio
Allnutt, Theo R
Powell, Wayne
author_sort Lee, David
collection PubMed
description BACKGROUND: The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR). Here we have applied the loop-mediated isothermal amplification (LAMP) method to amplify GMO-related DNA sequences, 'internal' commonly-used motifs for controlling transgene expression and event-specific (plant-transgene) junctions. RESULTS: We have tested the specificity and sensitivity of the technique for use in GMO studies. Results show that detection of 0.01% GMO in equivalent background DNA was possible and dilutions of template suggest that detection from single copies of the template may be possible using LAMP. CONCLUSION: This work shows that GMO detection can be carried out using LAMP for routine screening as well as for specific events detection. Moreover, the sensitivity and ability to amplify targets, even with a high background of DNA, here demonstrated, highlights the advantages of this isothermal amplification when applied for GMO detection.
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spelling pubmed-26564972009-03-17 Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences Lee, David La Mura, Maurizio Allnutt, Theo R Powell, Wayne BMC Biotechnol Research Article BACKGROUND: The most common method of GMO detection is based upon the amplification of GMO-specific DNA amplicons using the polymerase chain reaction (PCR). Here we have applied the loop-mediated isothermal amplification (LAMP) method to amplify GMO-related DNA sequences, 'internal' commonly-used motifs for controlling transgene expression and event-specific (plant-transgene) junctions. RESULTS: We have tested the specificity and sensitivity of the technique for use in GMO studies. Results show that detection of 0.01% GMO in equivalent background DNA was possible and dilutions of template suggest that detection from single copies of the template may be possible using LAMP. CONCLUSION: This work shows that GMO detection can be carried out using LAMP for routine screening as well as for specific events detection. Moreover, the sensitivity and ability to amplify targets, even with a high background of DNA, here demonstrated, highlights the advantages of this isothermal amplification when applied for GMO detection. BioMed Central 2009-02-02 /pmc/articles/PMC2656497/ /pubmed/19187544 http://dx.doi.org/10.1186/1472-6750-9-7 Text en Copyright © 2009 Lee et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Lee, David
La Mura, Maurizio
Allnutt, Theo R
Powell, Wayne
Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences
title Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences
title_full Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences
title_fullStr Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences
title_full_unstemmed Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences
title_short Detection of genetically modified organisms (GMOs) using isothermal amplification of target DNA sequences
title_sort detection of genetically modified organisms (gmos) using isothermal amplification of target dna sequences
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2656497/
https://www.ncbi.nlm.nih.gov/pubmed/19187544
http://dx.doi.org/10.1186/1472-6750-9-7
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