The SureFood® GMO SCREEN 4plex BAR/PAT/CryIAb/IAc/CTP2:CP4 EPSPS is a real-time PCR for the direct, qualitative detection and differentiation of following specific genetically modified DNA – sequences.
- Phosphinothricin-Acetyltransferase gene (BAR) from Streptomyces hygroscopius
- genetically engineered CryIAb-DNA-sequences and CryIAb/Ac-fusion gene sequences
- Phosphinothricin-Acetyltransferase gene (PAT) from Streptomyces viridiochromogenes
- the transition from CTP2 (Chloroplast-Transpeptide-signal sequence from Arabidopsis thaliana) to herbicide tolerance-gene CP4 EPSPS (5-Enolpyruvylshikimat-3-Phosphat Syntheses gene from Agrobacterium tumefaciens strain CP4)
The detections are according to the validated methods of the European Commission or the official collection of detection methods of §64 German food law.
The real-time PCR assay can be performed with commonly used real-time PCR instruments, equipped for detection of four fluorescence emissions at the channels FAM, VIC/HEX, ROX and Cy5 at the same time. The technical validation of instruments was performed on Agilent Mx3005P, Bio-Rad CFX96, Roche LightCycler® 480 II, Applied Biosystems 7500, Qiagen Rotor-Gene Q and R-Biopharm RIDA®CYCLER.
|Sample preparation||For DNA-preparation of raw material the use of SureFood® PREP Basic and for highly processed food and feed the use of SureFood® PREP Advanced is recommended. SureFood® PREP Add On is intended to be used for the extraction of DNA from raw materials as well as processed food and feed with sample weight of 2 g. It is used in conjunction with the SureFood® PREP Basic.|
|LOD (Detection limit)||The SureFood® GMO SCREEN 4plex BAR/PAT/CryIAb/IAc/CTP2:CP4 EPSPS real-time PCR has a limit of detection of < 5 DNA copies. This is equivalent to a limit of detection of <0.01 % based on the genetically modified organisms MON87751 Soya (CryIAb/IAc), DAS-81419-2 Soya (PAT), NK603 Corn (CTP2:CP4 EPSPS) and Bt176 Corn (BAR).|