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Naslov:Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR
Avtorji:ID Bogožalec Košir, Alexandra (Avtor)
ID Muller, Sabine (Avtor)
ID Žel, Jana (Avtor)
ID Milavec, Mojca (Avtor)
ID Mallory, Allison C. (Avtor)
ID Dobnik, David (Avtor)
Datoteke:URL URL - Izvorni URL, za dostop obiščite https://doi.org/10.3390/foods12224156
 
.pdf PDF - Izvorna datoteka, prenos (1,83 MB)
MD5: 330DC3B84B961FAC517B5F92875FD051
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo NIB - Nacionalni inštitut za biologijo
Povzetek:The proliferation of genetically modified organisms (GMOs) presents challenges to GMO testing laboratories and policymakers. Traditional methods, like quantitative real-time PCR (qPCR), face limitations in quantifying the increasing number of GMOs in a single sample. Digital PCR (dPCR), specifically multiplexing, offers a solution by enabling simultaneous quantification of multiple GMO targets. This study explores the use of the Naica six-color Crystal dPCR platform for quantifying five GM soybean lines within a single six-plex assay. Two four-color assays were also developed for added flexibility. These assays demonstrated high specificity, sensitivity (limit of detection or LOD < 25 copies per reaction) and precision (bias to an estimated copy number concentration <15%). Additionally, two approaches for the optimization of data analysis were implemented. By applying a limit-of-blank (LOB) correction, the limit of quantification (LOQ) and LOD could be more precisely determined. Pooling of reactions additionally lowered the LOD, with a two- to eight-fold increase in sensitivity. Real-life samples from routine testing were used to confirm the assays’ applicability for quantifying GM soybean lines in complex samples. This study showcases the potential of the six-color Crystal dPCR platform to revolutionize GMO testing, facilitating comprehensive analysis of GMOs in complex samples.
Ključne besede:digital PCR, dPCR, quantification, multiplexing, genetically modified organisms, 6-color system, virus diagnostics, virology
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Datum objave:01.11.2023
Leto izida:2023
Št. strani:str. [1]-17
Številčenje:iss. 22, art. no. ǂ4156
PID:20.500.12556/DiRROS-18583 Novo okno
UDK:578
ISSN pri članku:2304-8158
DOI:10.3390/foods12224156 Novo okno
COBISS.SI-ID:173360131 Novo okno
Opomba:Soavtorji: Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory and David Dobnik; Nasl. z nasl. zaslona; Opis vira z dne 23. 11. 2023;
Datum objave v DiRROS:29.03.2024
Število ogledov:94
Število prenosov:41
Metapodatki:XML RDF-CHPDL DC-XML DC-RDF
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Gradivo je del revije

Naslov:Foods
Skrajšan naslov:Foods
Založnik:MDPI
ISSN:2304-8158
COBISS.SI-ID:512252472 Novo okno

Gradivo je financirano iz projekta

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:P4-0165-2022
Naslov:Biotehnologija in sistemska biologija rastlin

Financer:Drugi - Drug financer ali več financerjev
Program financ.:European Union, European Regional Development Fund

Licence

Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

Jezik:Slovenski jezik
Ključne besede:digitalna PCR, digitalna verižna reakcija s polimerazo, kvantifikacija, multipleksiranje, gensko spremenjeni organizmi, 6-barvni sistem, diagnostika virusov, virologija


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