Sensitive and rapid detection of genetic modified soybean (Roundup Ready) by loop-mediated isothermal amplification.
Using the LAMP method, a highly specific and sensitive detection system for genetically modified soybean (Roundup Ready) was designed. In this detection system, a set of four primers was designed by targeting the exogenous 35S epsps gene. Target DNA was amplified and visualized on agarose gel within 45 min under isothermal conditions at 65 degrees C. Without gel electrophoresis, the LAMP amplicon was visualized directly in the reaction tube by the addition of SYBR Green I for naked-eye inspection. The detection sensitivity of LAMP was 10-fold higher than the nested PCR established in our laboratory. Moreover, the LAMP method was much quicker, taking only 70 min, as compared with 300 min for nested PCR to complete the analysis of the GM soybean. Compared with traditional PCR approaches, the LAMP procedure is faster and more sensitive, and there is no need for a special PCR machine or electrophoresis equipment. Hence, this method can be a very useful tool for GMO detection and is particularly convenient for fast screening.
Liu
Mei
M
R&D Center of Marine Biotechnology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.
Luo
Yan
Y
Tao
Ran
R
He
Ru
R
Jiang
Keyong
K
Wang
Baojie
B
Wang
Lei
L
D017931:DNA Primers / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics; D005998:Glycine / Q000031:analogs & derivatives*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D013025:Glycine max / Q000235:genetics*; D013997:Time Factors; D000097797:Glyphosate
D017931:DNA Primers; D005998:Glycine; D000097797:Glyphosate