20.500.12556/DiRROS-20132 Critical assessment of digital PCR for the detection and quantification of genetically modified organisms 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. digital PCR droplet digital PCR chip-based digital PCR genetically modified organisms quantification true false true Angleški jezik Ni določen Neznano 2024-08-06 09:28:03 2024-08-06 09:28:03 2024-08-08 03:56:46 0000-00-00 00:00:00 2018 0 0 str. 4039-4050 iss. 17 Vol. 410 2018 0000-00-00 Zaloznikova NiDoloceno NiDoloceno 0000-00-00 0000-00-00 2018-03-24 604.6 1618-2642 10.1007/s00216-018-1010-1 4657231 http://dx.doi.org/10.1007/s00216-018-1010-1 1 a9e76a9a-53b4-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=28100 RAZ_Demeke_Tigst_2018.pdf RAZ_Demeke_Tigst_2018.pdf 1 6107CF7EC311D518B13FE50664B6382A 2ece70d696d3f8e7cf09b725c8b41f890d49c7ed805f01d85e2bbd1b04e1d51c e67b1a24-53b4-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=28101 Nacionalni inštitut za biologijo 0 0 0