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A high-throughput analytical method for multiple DNA targets based on microdroplet PCR coupled with DGGE

文献类型： 外文期刊

作者： Zhao, Binan 4 ; Zhao, Xiao 3 ; Yang, Dan 1 ; Pu, Xinyi 1 ; Xu, Yan 1 ; Zhang, Xiaoxia 1 ; Wu, Wenjing 1 ; Zhang, Wanjing 1 ; Sun, Chuanwen 1 ; Zhang, Qi 1 ; Zhao, Kai 2 ;

作者机构： 1.Shanghai Normal Univ, Coll Life Sci, Shanghai 200234, Peoples R China

2.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai 201106, Peoples R China

3.Ningxia Univ, Coll Life Sci, Ningxia 750021, Peoples R China

4.Tongji Univ, Shanghai Peoples Hosp 10, Sch Med, Shanghai 200092, Peoples R China

关键词： Nucleic acid detection; Microdroplet PCR; DGGE; MPDG; Transgenic detection; GM maize

期刊名称：JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS （ 影响因子：3.571； 五年影响因子：3.552 ）

ISSN： 0731-7085

年卷期： 2022 年 216 卷

页码：

收录情况： SCI

摘要： We developed a novel approach to analyze multiple DNA targets based on microdroplet PCR coupled with denaturing gradient gel electrophoresis (MPDG) to achieve high-throughput detection of biological samples. The target DNAs were preamplified using specific primers. Subsequently, the preamplified products were separated into individual microreactors for parallel amplification with high efficiency, avoiding the interference of different primers and templates, and preventing inconsistent amplification efficiency and non-specific amplification. The final products were analyzed using denaturing gradient gel electrophoresis (DGGE). Using genetically modified (GM) maize as samples, the MPDG method could be used for the simultaneous detection of three DNA targets with an absolute limit of detection of 0.5% (w/w), with no cross reaction with other non-GM samples. The simulated sample assay of MPDG suggested that the method is suitable for practical application. The MPDG approach, with high sensitivity and specificity, could play a crucial role in the field of nucleic acid detection.

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