Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 410, Issue 17, Pages 4039-4050Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-018-1010-1
Keywords
Digital PCR; Droplet digital PCR; Chip-based digital PCR; Genetically modified organisms; Quantification
Funding
- European Union [613908]
- Slovenian Research Agency [P4-0165, 1000-15-0105]
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The number of genetically modified organisms (GMOs) on the market is steadily increasing. Because of regulation of cultivation and trade of GMOs in several countries, there is pressure for their accurate detection and quantification. Today, DNA-based approaches are more popular for this purpose than protein-based methods, and real-time quantitative PCR (qPCR) is still the gold standard in GMO analytics. However, digital PCR (dPCR) offers several advantages over qPCR, making this new technique appealing also for GMO analysis. This critical review focuses on the use of dPCR for the purpose of GMO quantification and addresses parameters which are important for achieving accurate and reliable results, such as the quality and purity of DNA and reaction optimization. Three critical factors are explored and discussed in more depth: correct classification of partitions as positive, correctly determined partition volume, and dilution factor. This review could serve as a guide for all laboratories implementing dPCR. Most of the parameters discussed are applicable to fields other than purely GMO testing.
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