Statistical Evaluation of Real-Time PCR Protocols Applied to Quantify Genetically Modified Maize

Statistical Evaluation of Real-Time PCR Protocols Applied to Quantify Genetically Modified Maize Real-time PCR (RTi-PCR) is the technique of choice for event-specific quantification of genetically modified organisms (GMOs) by determining the amount of event with respect to a species-specific reference gene. Reference genes can be amplified from the genome extracted from Certified Reference Materials (CRMs) or from ad hoc designed plasmids. In the present study, we statistically evaluate the performance of RTi-PCR protocols for GM maize MON810 event by using both genomic DNA from conventional CRMs and a plasmid containing sequences representative of four maize species-specific reference genes. The significance of simple and interaction effects of several variables included in the experimental design on DNA quantification methods and RTi-PCR were evaluated and discussed. Statistically significant differences on Ct values may have an impact on the GMOs quantification and consequently on the compliance of GM quantification-established legal thresholds. Our results confirm the reliability of the plasmid as alternative calibrant for the calculation of GMOs copy number. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Food Analytical Methods Springer Journals

Statistical Evaluation of Real-Time PCR Protocols Applied to Quantify Genetically Modified Maize

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Publisher
Springer Journals
Copyright
Copyright © 2010 by Springer Science+Business Media, LLC
Subject
Chemistry; Analytical Chemistry ; Microbiology ; Chemistry/Food Science, general ; Food Science
ISSN
1936-9751
eISSN
1936-976X
DOI
10.1007/s12161-010-9135-7
Publisher site
See Article on Publisher Site

Abstract

Real-time PCR (RTi-PCR) is the technique of choice for event-specific quantification of genetically modified organisms (GMOs) by determining the amount of event with respect to a species-specific reference gene. Reference genes can be amplified from the genome extracted from Certified Reference Materials (CRMs) or from ad hoc designed plasmids. In the present study, we statistically evaluate the performance of RTi-PCR protocols for GM maize MON810 event by using both genomic DNA from conventional CRMs and a plasmid containing sequences representative of four maize species-specific reference genes. The significance of simple and interaction effects of several variables included in the experimental design on DNA quantification methods and RTi-PCR were evaluated and discussed. Statistically significant differences on Ct values may have an impact on the GMOs quantification and consequently on the compliance of GM quantification-established legal thresholds. Our results confirm the reliability of the plasmid as alternative calibrant for the calculation of GMOs copy number.

Journal

Food Analytical MethodsSpringer Journals

Published: Mar 30, 2010

References

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