Research On PCR Chip For Detection Of Genetically Modified Maize

Transgenic technology is a hot topic in the field of agriculture.The research of transgenic maize is one of the orientation in transgenic technology,has been wildly studied over the past several year,and great economic benefits have been brought.However,a series of problems have arisen,such as the impact on physical health,the potential hazards to ecological environment and so on.It was very important and urgent to strengthen the detection of transgenic products to ensure the implementation of identity management system and safeguard people’s right to know the truth.So finding high sensitivity and high accuracy methods has become a research hot spot object.In this study,we summarized the principles and the features of various detection methods of genetically modified maize,and some problems are discussed when these methods are used for detecting transgenic products.In order to extract high quality genome DNA from maize,six methods were adopted to optimize genome DNA extraction in this research.Finally,we found the most efficient method for genome DNA extraction from maize,and the method would be of great help to the real-time PCR method.This experiment of the real-time PCR selected 13 GM maize lines(BT11、BT176、GA21、MON810、MON863、MON89034、NK603、MON88017、MIR604、MIR162、DP98140、BT10、T25),and analysed the sequences of transgenic and maize endogenous reference gene ZEIN by using NCBI databases,13 primer pairs and probes with high specificity were designed using Primer5.0 and DNAMAN.Through the experiment of the real-time PCR,the amplified condition were confirmed,and sensitivity and specificity of primer pairs and probes were proved.The study constructed a PCR chip for detecting several types of genetically modified maize in corn and their productions.The PCR chip is improved on the basis of PCR microtiter plate and combined the merits of microfluidics.The volume of one well is reduced to 2.5L,and 384 wells in a chip.In the experiment,seven kinds of genetically modified maize(BT10、BT11、BT176、GA21、MON863、MON89034、T25)and one non-GM maize were used to confirm the amplified condition,sensitivity and specificity of PCR chip.The results confirmed that this PCR chip has good sensitivity and specificity,the detection limit could reach 0.1%.According to the specificity and sensitivity test,it shows that the PCR chip is good enough to be applied in the identification of genetic modified maize and its products.