20.500.12556/DiRROS-19787 Multiplex quantification of four DNA targets in one reaction with Bio-Rad droplet digital PCR system for GMO detection The advantages of the digital PCR technology are already well documented until now. One way to achieve better cost efficiency of the technique is to use it in a multiplexing strategy. Droplet digital PCR platforms, which include two fluorescence filters, support at least duplex reactions and with some developments and optimization higher multiplexing is possible. The present study not only shows a development of multiplex assays in droplet digital PCR, but also presents a first thorough evaluation of several parameters in such multiplex digital PCR. Two 4-plex assays were developed for quantification of 8 different DNA targets (7 genetically modified maize events and maize endogene). Per assay, two of the targets were labelled with one fluorophore and two with another. As current analysis software does not support analysis of more than duplex, a new R- and Shiny-based web application analysis tool (http://bit.ly/ddPCRmulti) was developed that automates the analysis of 4-plex results. In conclusion, the two developed multiplex assays are suitable for quantification of GMO maize events and the same approach can be used in any other field with a need for accurate and reliable quantification of multiple DNA targets. digital PCR DNA targets GMO detection true false true Angleški jezik Ni določen Neznano 2024-07-25 04:13:41 2024-07-25 17:55:10 2024-07-26 04:28:26 0000-00-00 00:00:00 2016 0 0 Nasl. z nasl. zaslona; Opis vira z dne 18. 10. 2016; str. 1-9 Vol. 6 2016 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2016-10-14 577.2 2045-2322 10.1038/srep35451 4055119 RAZ_Dobnik_David_2016.pdf RAZ_Dobnik_David_2016.pdf 1 2C48F9F0225704E5DEACF52741B9A2CD b8dd424b0dc2e576c555dc322e7b243ec9273f051afa12dbe094678163a51d7c ae8b257a-4a8d-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27551 https://doi.org/10.1038/srep35451 0 9c1229a5-4a8d-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27550 Nacionalni inštitut za biologijo 0 0 0