New plasmid calibrators for geminivirus-resistant (EMB-PV051-1 event) common bean (Phaseolus vulgaris L.) quantitation using simplex and duplex qPCR.
2018
VENTURELLI, G. L. | SILVA, K. J. DA | TREML, D. | NAVAS, P. B. | VARGAS, M. O. | BISCHOFF, J. L. | FARIA, J. C. DE | ARISI, A. C. M. | GUSTAVO LUIZ VENTURELLI, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC; KELLY JUSTIN DA SILVA, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC; DIANA TREML, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC; PAOLA BEATRIZ NAVAS, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC; MARYELLA OSÓRIO VARGAS, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC; JOANA LAURA BISCHOFF, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC; JOSIAS CORREA DE FARIA, CNPAF; ANA CAROLINA MAISONNAVE ARISI, UNIVERSIDADE FEDERAL DE SANTA CATARINA, Florianópolis-SC.
The geminivirus-resistant common bean, Embrapa 5.1, was the first commercial genetically modified (GM) event developed in Latin America by the Brazilian Agricultural Research Corp.. Therein novel standard reference plasmids were constructed for species-specific (pLEC) and event-specific (pFGM) qualitative and quantitative detection of Embrapa 5.1 GM common bean. To establish these plasmids as certified reference materials (CRM) for Embrapa 5.1 GM common bean, two DNA extraction protocols, simplex and duplex qPCR using two different operators (experimenters) were tested. The efficiency values ranged from 92% to 110% for the simplex and duplex reactions considering both operators. The limit of detection was enough to detect at least 0.1% GM content. These plasmids are suitable to be used as CRM for Embrapa 5.1 GM common bean. They will be useful for survey of food labels for compliance with legislation about GMO content in Brazil and in other countries where GM common bean is not yet approved for commercialization.
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