Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 55, Issue 9, Pages 3249-3257Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jf062931l
Keywords
feed; food; genetically modified organism; GM; measurement uncertainty; extraction method; DNA; PCR
Funding
- Ministry of Education, Science & Technology (MoST), Republic of Korea [07-2404-022] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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An international CCQM-P60 pilot study involving eight national metrological institutes was organized to investigate if the quantification of genetically modified (GM) corn powder by real-time PCR was affected by the DNA extraction method applied. Four commonly used extraction methods were compared for the extraction of DNA from a GM Bt176 corn powder. The CTAB-based method yielded the highest DNA template quantity and quality. A difference in the 260 nm/230 nm absorbance ratio was observed among the different extraction methods. Real-time amplification of sequences specific for endogenous genes zein and hmg as well as transgenic sequences within the cryIA(b) gene and a fragment covering the junction between the transformed DNA and the plant genome were used to determine the GM percentage. The detection of the transgenic gene was affected by the quantity and quality of template used for the PCR reaction. The Bt176 percentages measured on diluted or purified templates were statistically different depending on the extraction method applied.
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