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| Validation Certified Value of Multi-targets Plasmid Reference Materials by Three Digital PCR Platforms |
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Abstract Abstract Digital PCR is a new absolute quantitative method that does not rely on the external standard curves, and it shows broad application prospects in quantified detection of genetically modified organisms (GMOs), evaluation the exogenous gene copy number of GMOs and validation the certified values of GM reference materials. In this study, three digital PCR platforms including chamber digital PCR (cdPCR), droplet digital PCR (ddPCR) and 3D digital PCR (3D-dPCR) were used to quantify the copy number of exogenous fragments and endogenous gene phosphoenolpyruvate carboxylase (PEP) of GM canola (Brassica campestris) multi-targets plasmid reference molecule, and the certified value were determined by the mean copy number ratios of exo-gene/endo-gene. The results indicated that the certified values from three dPCR technology were close to the true value of 1 (copies/copies) and the relative standard deviations were (RSD) below 25%, The three dPCR all had stable reaction systems and good repeatability and showed high accuracy in value determination of plasmid DNA reference materials. Meanwhile, the plasmid value determination methods based on cdPCR, ddPCR and 3D-dPCR platforms provide a new model for validation of other reference materials' certified values.
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Received: 20 January 2017
Published: 06 August 2017
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