title: Development of plasmid DNA reference material for the quantification of genetically modified common bean embrapa 5.1.
abstract: The genetically modified (GM) common bean Embrapa 5.1 was recently approved for commercialization. The reliable detection and quantification of GM organisms is strongly dependent on validated methods as well as calibration systems. This work presents the development of a calibrant plasmid for Embrapa 5.1 common bean detection. The reaction parameters were determined and compared for both the plasmid DNA (pDNA) and the genomic DNA (gDNA). PCR efficiencies for pDNA were 81% for the construction-specific assays and 76% for the taxon-specific assay, whereas for gDNA efficiencies were 94 and 93%, respectively. The limits of detection (LOD) in both qPCR assays were 10(2) and 10(3) copies of gDNA and pDNA per PCR reaction, respectively. This is sufficient to detect 0.067 and 0.67% of GM common bean in 100 ng of DNA, respectively, which is in agreement with detecting the 1% GM content required by the Brazilian legislation.
authors.affiliation: Departamento de Ciência e Tecnologia de Alimentos, Centro de Ciências Agrárias, Universidade Federal de Santa Catarina, Rod. Admar Gonzaga 1346, 88034-001 Florianópolis, SC, Brazil.
authors.forename: Fábio C A
authors.initials: FC
authors.lastname: Brod
authors.forename: Andréia Z
authors.initials: AZ
authors.lastname: Dinon
authors.forename: Deise J
authors.initials: DJ
authors.lastname: Kolling
authors.forename: Josias C
authors.initials: JC
authors.lastname: Faria
authors.forename: Ana C M
authors.initials: AC
authors.lastname: Arisi
mesh_terms: D001938:Brazil; D002138:Calibration; D018744:DNA, Plant / Q000032:analysis* / Q000235:genetics*; D005818:Genetic Engineering / Q000331:legislation & jurisprudence; D057230:Limit of Detection; D027805:Phaseolus / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards
chemical_list: D018744:DNA, Plant