Development and application of a quantitative loop-mediated isothermal amplification method for detecting genetically modified maize MON863

Autor:	Sicong, Huang, Yuancong, Xu, Xinghua, Yan, Ying, Shang, Pengyu, Zhu, Wenying, Tian, Wentao, Xu
Rok vydání:	2013
Předmět:	DNA Plant Food Genetically Modified Temperature Reproducibility of Results Haploidy Genes Plant Plants Genetically Modified Polymerase Chain Reaction Zea mays Betaine Humans Magnesium Nucleic Acid Amplification Techniques Food Analysis Genome Plant DNA Primers
Zdroj:	Journal of the science of food and agriculture. 95(2)
ISSN:	1097-0010
Popis:	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.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.The results of this study demonstrate that this new SYBR Green I-based quantitative LAMP assay system is reliable, sensitive and accurate.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::5322bddd8733f0d408d6267414babfa1 https://pubmed.ncbi.nlm.nih.gov/24771525 Zobrazit plný text záznamu Plný text