A novel method was proposed for the detection of loop mediated isothermal amplification(LAMP) products of Escherichia coli(E-coli)based on self-built capillary electrophoresis(CE)with lamp induced fluorescence system.Four primers were designed for the target DNA,and the sizes of the LAMP products of E-coli corresponding to the primers of F3,B3,FIP and BIP were 240 bp.The best result was achieved with the sieving polymer of 0.5% HEC(1 300 K),an electric field strength of 100 V/cm and an injection time of 1.0 s at the electric field strength of 100 V/cm.The migration time of DNA from 100 bp to 500 bp is linearly related with DNA size,and the correlation coefficient is 0.996.The LAMP products of E-coli were resolved under the same electrophoretic conditions.The sizes of the LAMP products were calculated based on the linear relationship,and the positive products was distinguished from the false-positive products.The results demonstrated that CE could not only rapidly separate the LAMP products of E-coli within 15 min,but also distinguish the positive from false-positive products.The proposed method was successfully applied in the separation of the LAMP products of AB0174 E-coli.Therefore,it is suitable for the rapid detection of LAMP products of DNA.
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School of Environmental and Pharmaceutical Engineering,Taizhou Institute of Science and Technology,Nanjing University of Science and Technology
Jiangsu Key Laboratory of Environmental Science and Engineering
School of Environmental Science and Engineering,Suzhou University of Science and Technology