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Title:Critical assessment of digital PCR for the detection and quantification of genetically modified organisms
Authors:ID Demeke, Tigst (Author)
ID Dobnik, David (Author)
Files:URL URL - Source URL, visit http://dx.doi.org/10.1007/s00216-018-1010-1
 
.pdf PDF - Presentation file, download (978,27 KB)
MD5: 6107CF7EC311D518B13FE50664B6382A
 
Language:English
Typology:1.02 - Review Article
Organization:Logo NIB - National Institute of Biology
Abstract: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.
Keywords:digital PCR, droplet digital PCR, chip-based digital PCR, genetically modified organisms, quantification
Publication version:Version of Record
Publication date:24.03.2018
Year of publishing:2018
Number of pages:str. 4039-4050
Numbering:Vol. 410, iss. 17
PID:20.500.12556/DiRROS-20132 New window
UDC:604.6
ISSN on article:1618-2642
DOI:10.1007/s00216-018-1010-1 New window
COBISS.SI-ID:4657231 New window
Publication date in DiRROS:06.08.2024
Views:14
Downloads:4
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Record is a part of a journal

Title:Analytical and bioanalytical chemistry
Shortened title:Anal. bioanal. chem.
Publisher:Springer
ISSN:1618-2642
COBISS.SI-ID:6966294 New window

Document is financed by a project

Funder:EC - European Commission
Project number:613908
Name:Development of Cost efficient Advanced DNA-based methods for specific Traceability issues and High Level On-site applicatioNs
Acronym:DECATHLON

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0165-2015
Name:Biotehnologija in sistemska biologija rastlin

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:1000-15-0105

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