Volume 9, Number 2, p.p. 73–76
Optimization of DNA-based screening methods for genetically modified organisms
Tamara Kutateladze,1 Marina Karseladze,1 Inga Gabriadze,1 and Nelly Datukishvili1,2
1
Institute of Molecular Biology and Biological Physics, 12 Gotua St, 0160 Tbilisi, Georgia
2
Ilia Chavchavadze State University, 32 Chavchavadze Ave, 0179 Tbilisi, Georgia
The worldwide distribution of genetically engineered plants has generated an urgent need for reliable and efficient methods of monitoring genetically modified organisms (GMOs) in each country. In this study, certified reference materials in the form of dried powders containing 0–5% Roundup Ready soybean and maize MON-810 were used for optimization and validation of GMO screening methods. Genomic DNAs were isolated using a Qiagen DNeasy plant minikit. The high amplification quality of extracted DNAs was revealed by a plant-specific polymerase chain reaction (PCR) suitable for the detection of chloroplast genome conserved sequences. The conventional end-point polymerase chain reactions specific to the 35S promoter from cauliflower mosaic virus and the NOS terminator from Agrobacterium tumefaciens confirmed the presence of transgenic material in the GMO-containing reference materials with high specificity and a sensitivity of 0.1% GMO, while negative water and non-GM samples did not generate any amplification signal. The results obtained indicate that the DNA-based analytical procedure described in this paper allows a reliable and sensitive qualitative detection of GMOs that corresponds to European regulatory requirements.
Keywords:
DNA analysis, NOS terminator, qualitative detection, transgenic products, 35S promoter