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Development and application of a quantitative loop‐mediated isothermal amplification method for detecting genetically modified maize MON863
Author(s) -
Huang Sicong,
Xu Yuancong,
Yan Xinghua,
Shang Ying,
Zhu Pengyu,
Tian Wenying,
Xu Wentao
Publication year - 2014
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.6707
Subject(s) - loop mediated isothermal amplification , betaine , sybr green i , genetically modified organism , quantitative analysis (chemistry) , microbiology and biotechnology , genomic dna , real time polymerase chain reaction , chemistry , isothermal process , dna , biology , chromatography , gene , biochemistry , thermodynamics , physics
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. CONCLUSION The results of this study demonstrate that this new SYBR Green I‐based quantitative LAMP assay system is reliable, sensitive and accurate. © 2014 Society of Chemical Industry
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