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SanJuan-Badillo,Andrea; Galvez,Amanda; Plasencia,Javier; Quirasco,Maricarmen. |
In Mexico, regulations for growing genetically modified (GM) maize (Zea mays L.) plants have been enforced in order to prevent gene flow to native landraces and wild relatives. Field surveys are necessary and Mexican government agencies have the mandate to perform them. Because of their specificity, PCR-based methods are suitable for field monitoring of GM organisms but it is necessary to assess their performance, since they are greatly influenced by the DNA preparation quality inherent to the extraction method. In this study, genomic DNA was extracted from various maize tissues (e.g. pollen, leaves, spikelets and grains) using five different commercial purification protocols. DNA quality was analyzed spectrophotometrically, by gel electrophoresis, and as... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: PCR inhibition; GM maize; Variability of DNA yield; DNA quantification; GMO detection. |
Ano: 2014 |
URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952014000100002 |
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Acatzi,Abraham; Galvez,Amanda; Plasencia,Javier; Quirasco,Maricarmen. |
Because of their specificity and high throughput, real-time PCR-based methods are suitable for the monitoring of genetically modified (GM) organisms at field level or in the grain trade, in order to comply with federal regulations on biosafety. In the first part of this study, DNA extracted from different maize (Zea mays L.) tissues using several commercial purification protocols available in Mexico were evaluated in terms of DNA quality as substrates for end-point PCR. In this second part, DNA preparations obtained from grain, by means of the same commercial protocols, were tested in quantitative PCR (qPCR), using TaqMan and SYBR Green protocols. Linear dynamic range, amplification efficiency and method accuracy were assessed using recommended qPCR... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Quantitative PCR; PCR inhibition; GM maize; Genetically modified organism (GMO); DNA quality. |
Ano: 2014 |
URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952014000100003 |
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