Development and application of a quantitative loop-mediated isothermal amplification method for detecting genetically modified maize MON863.
BACKGROUND
A SYBR Green I-based quantitative loop-mediated isothermal amplification (LAMP) assay was developed for the rapid detection of genetically modified maize MON863. A set of primers was designed based on the integration region of the Cry3Bb1 and tahsp17 genes.


RESULTS
The qualitative and quantitative reaction conditions (dNTPs, betaine, primers, Mg(2+), Bst polymerase, temperature, reaction time) were optimized. The concentrations of Mg(2+) and betaine were found to be important to the LAMP assay. The detection limits of both qualitative and quantitative LAMP for MON863 were as low as 4 haploid genomic DNA, and the LAMP reactions can be completed within 1 h at an isothermal temperature of 65 °C.


CONCLUSIONS
The results of this study demonstrate that this new SYBR Green I-based quantitative LAMP assay system is reliable, sensitive and accurate.
High School attached to Tsinghua University, Beijing, 100084, China.
Sicong
S
Huang
Yuancong
Y
Xu
Xinghua
X
Yan
Ying
Y
Shang
Pengyu
P
Zhu
Wenying
W
Tian
Wentao
W
Xu
D001622:Betaine / Q000032:analysis; D017931:DNA Primers; D018744:DNA, Plant / Q000032:analysis*; D005504:Food Analysis / Q000379:methods*; D030841:Food, Genetically Modified; D017343:Genes, Plant; D018745:Genome, Plant; D006238:Haploidy; D006801:Humans; D008274:Magnesium / Q000032:analysis; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D030821:Plants, Genetically Modified / Q000737:chemistry; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013696:Temperature; D003313:Zea mays / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D018744:DNA, Plant; D001622:Betaine; D008274:Magnesium
Cry3Bb1; genetically modified maize; loop-mediated isothermal amplification; quantitative