Toward Metrological Traceability for DNA Fragment Ratios in GM Quantification. 2. Systematic Study of Parameters Influencing the Quantitative Determination of MON 810 Corn by Real-Time PCR

This paper is part of a set of three papers investigating metrological traceability of the quantification of DNA fragments as, for instance, used for quantification of genetic modifications. This paper evaluates the possible impact of several factors on results of real-time Polymerase Chain Reaction...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2007-05, Vol.55 (9), p.3258-3267
Main Authors: Charels, Diana, Broeders, Sylvia, Corbisier, Philippe, Trapmann, Stefanie, Schimmel, Heinz, Linsinger, Thomas, Emons, Hendrik
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
Bacterial Toxins - genetics
Biological and medical sciences
Cereal and baking product industries
corn
DNA fragment ratios
DNA, Plant - analysis
Endotoxins - genetics
extraction
feeds
Food engineering
Food industries
Food, Genetically Modified
Fundamental and applied biological sciences. Psychology
General aspects
genetically modified foods
Hemolysin Proteins - genetics
HSP70 Heat-Shock Proteins - genetics
MON 810 corn
particle size
Plants, Genetically Modified - genetics
polymerase chain reaction
Polymerase Chain Reaction - methods
powders
quantitative analysis
real time polymerase chain reaction
recombinant DNA
Seeds - genetics
traceability
transgenes
Zea mays - genetics
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Summary:This paper is part of a set of three papers investigating metrological traceability of the quantification of DNA fragments as, for instance, used for quantification of genetic modifications. This paper evaluates the possible impact of several factors on results of real-time Polymerase Chain Reaction (PCR) measurements. It was found that the particle size of the powder samples does not have an influence, whereas the nature of the calibrant (plasmidic or genomic DNA) has a significant effect. Moreover, two real-time PCR detection methods (construct-specific and event-specific) for MON 810 corn were compared. The results obtained in a specifically designed interlaboratory study revealed a significant influence of the DNA extraction method on measurement results when the MON 810 construct-specific real-time PCR detection method was applied. Statistical analyses confirmed the importance of validating DNA extraction methods in conjunction with real-time PCR methods. Keywords: Genetically modified organism; GMO; DNA; extraction; real-time PCR; PCR efficiency; DNA copy number ratio
ISSN:0021-8561
1520-5118
DOI:10.1021/jf062932d