2026-04-14T03:49:40Zhttps://recercat.cat/oai/request

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

00925njm 22002777a 4500 dc Nadal i Matamala, Anna author Esteve Nuez, Teresa author Pla i de Solà-Morales, Maria author 2009-05-01 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 http://hdl.handle.net/10256/25939 Reacció en cadena de la polimerasa Plantes transgèniques Cotó Polymerase chain reaction Transgenic plants Cotton Multiplex Polymerase Chain Reaction-Capillary Gel Electrophoresis: A Promising Tool for GMO ScreeningAssay for Simultaneous Detection of Five Genetically Modified Cotton Events and Species