2026-04-14T08:58:54Zhttps://recercat.cat/oai/request

oai:recercat.cat:10256/259392025-01-15T23:06:20Zcom_2072_452955com_2072_2054col_2072_453062

Multiplex Polymerase Chain Reaction-Capillary Gel Electrophoresis: A Promising Tool for GMO ScreeningAssay for Simultaneous Detection of Five Genetically Modified Cotton Events and Species Nadal i Matamala, Anna Esteve Nuez, Teresa Pla i de Solà-Morales, Maria Reacció en cadena de la polimerasa Plantes transgèniques Cotó Polymerase chain reaction Transgenic plants Cotton A multiplex polymerase chain reaction assay coupled to capillary gel electrophoresis for amplicon identification by size and color (multiplex PCR-CGE-SC) was developed for simultaneous detection of cotton species and 5 events of genetically modified (GM) cotton. Validated real-time-PCR reactions targeting Bollgard, Bollgard II, Roundup Ready, 3006-210-23, and 281-24-236 junction sequences, and the cotton reference gene acp1 were adapted to detect more than half of the European Union-approved individual or stacked GM cotton events in one reaction. The assay was fully specific (<1.7% of false classification rate), with limit of detection values of 0.1% for each event, which were also achieved with simulated mixtures at different relative percentages of targets. The assay was further combined with a second multiplex PCR-CGE-SC assay to allow simultaneous detection of 6 cotton and 5 maize targets (two endogenous genes and 9 GM events) in two multiplex PCRs and a single CGE, making the approach more economic. Besides allowing simultaneous detection of many targets with adequate specificity and sensitivity, the multiplex PCR-CGE-SC approach has high throughput and automation capabilities, while keeping a very simple protocol, e.g., amplification and labeling in one step. Thus, it is an easy and inexpensive tool for initial screening, to be complemented with quantitative assays if necessary 2009-05-01 info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion peer-reviewed http://hdl.handle.net/10256/25939 19610365 http://hdl.handle.net/10256/25939 eng info:eu-repo/semantics/altIdentifier/doi/10.1093/jaoac/92.3.765 info:eu-repo/semantics/altIdentifier/issn/1060-3271 info:eu-repo/semantics/altIdentifier/eissn/1944-7922 Tots els drets reservats info:eu-repo/semantics/openAccess 8 p. application/pdf Oxford University Press © Journal of AOAC International, 2009, vol. 92, núm. 3, p. 765-772 Articles publicats (D-EQATA) Nadal i Matamala, Anna Esteve Nuez, Teresa Pla i de Solà-Morales, Maria 2009 Multiplex Polymerase Chain Reaction-Capillary Gel Electrophoresis: A Promising Tool for GMO ScreeningAssay for Simultaneous Detection of Five Genetically Modified Cotton Events and Species Journal of AOAC International 92 3 765 772