| Autor: |
Pardigol, Andreas, Guillet, Stéphanie, Pöpping, Bert |
| Zdroj: |
European Food Research and Technology; 20030501, Vol. 216 Issue: 5 p412-420, 9p |
| Abstrakt: |
Abstract Determination of the genetically modified organism (GMO) content in foods and feeds by means of real-time polymerase chain reaction (PCR) requires appropriate calibration standards. For this purpose, commercial certified reference materials calibrants (CRMCs) are commonly used. However, quantitative results depend on the actual GMO content of the standards, which may vary from lot to lot. Furthermore, commercial CRMCs are available in the range of 0%-5% only, thus limiting the quantification working range. More recently, recombinant plasmid standards have been used as an alternative to commercial CRMCs. Their production, however, necessitates cloning and microbiology facilities. Here we describe an easy and cost-effective procedure for construction of hybrid amplicon standards containing both transgene and reference gene targets in a tandem orientation on the same molecule. These standards can be rapidly produced in abundant amounts without any equipment for cloning and culturing of recombinant strains. This approach has proved accurate and precise and exhibits major advantages over genomic DNA or plasmids. In this paper we present a validation study for quantitative analysis of Roundup Ready soybean and the 35S-CaMV promoter in genetically modified maize and soybean. |
| Databáze: |
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