| Title: | Inter-laboratory analysis of selected genetically modified plant reference materials with digital PCR |
|---|
| Authors: | ID Dobnik, David (Author)
ID Demšar, Tina (Author)
ID Huber, Ingrid (Author)
ID Gerdes, Lars (Author)
ID Broeders, Sylvia (Author)
ID Roosens, Nancy (Author)
ID Debode, Frédéric (Author)
ID Berben, Gilbert (Author)
ID Žel, Jana (Author) |
|---|
| Files: |  URL - Source URL, visit http://dx.doi.org/10.1007/s00216-017-0711-1
 PDF - Presentation file, download (466,94 KB)
MD5: 8CFF6CC0DAAB4C3D22938003F3295C09
|
|---|
| Language: | English |
|---|
| Typology: | 1.01 - Original Scientific Article |
|---|
| Organization: |  NIB - National Institute of Biology
|
|---|
| Abstract: | Digital PCR (dPCR), as a new technology in the field of genetically modified (GM) organism (GMO) testing, enables determination of absolute target copy numbers. The purpose of our study was to test the transferability of methods designed for quantitative PCR (qPCR) to dPCR and to carry out an inter-laboratory comparison of the performance of two different dPCR platforms when determining the absolute GM copy numbers and GM copy number ratio in reference materials certified for GM content in mass fraction. Overall results in terms of measured GM% were within acceptable variation limits for both tested dPCR systems. However, the determined absolute copy numbers for individual genes or events showed higher variability between laboratories in one third of the cases, most possibly due to variability in the technical work, droplet size variability, and analysis of the raw data. GMO quantification with dPCR and qPCR was comparable. As methods originally designed for qPCR performed well in dPCR systems, already validated qPCR assays can most generally be used for dPCR technology with the purpose of GMO detection. |
|---|
| Keywords: | digital PCR, droplet digital PCR, absolute quantification, reference materials, GMO quantification |
|---|
| Publication status: | Published |
|---|
| Publication version: | Version of Record |
|---|
| Publication date: | 01.01.2018 |
|---|
| Year of publishing: | 2018 |
|---|
| Number of pages: | str. 211-221 |
|---|
| Numbering: | Vol. 410, iss. 1 |
|---|
| PID: | 20.500.12556/DiRROS-19632  |
|---|
| UDC: | 604.6 |
|---|
| ISSN on article: | 1618-2642 |
|---|
| DOI: | 10.1007/s00216-017-0711-1 |
|---|
| COBISS.SI-ID: | 4500303 |
|---|
| Publication date in DiRROS: | 24.07.2024 |
|---|
| Views: | 433 |
|---|
| Downloads: | 169 |
|---|
| Metadata: |    |
|---|
|
:
|
Copy citation |
|---|
| | | | Share: |  |
|---|
Hover the mouse pointer over a document title to show the abstract or click
on the title to get all document metadata. |
