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Abstract With the development of genetic modified plants and products, the urgent need is the standard reference materials for genetic modified detection. This work is about genetic modified maize NK603 standard plasmid construction and application. Genetic modified (GM) maize (Zea mays) line NK603 event specific product and endogenous gene zSSIIb product were amplified and ligated to T vector to constructed plasmid pNK603. And the pNK603 plasmid was successfully proofed by PCR amplification, digestion and sequencing. Results showed that the pNK603 plasmid was very specific for GM maize NK603 since there were no any other PCR product amplified using different primers except the specific main bands. Three different level of GM matrix samples were detected by quantitative PCR when pNK603 plasmid DNA and GM NK603 genomic DNA were used as quantification standards, respectively. The result showed that there was no statistic significance between the two different standards. T test showed the PCR efficiency, slop and intercept of the two standards for NK603 and endogenous gene respectively were not significantly different. The results indicated that pNK603 plasmid can be used as a standard for GM maize NK603 detection.
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Received: 02 August 2010
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