MPIC: A High-Throughput Analytical Method for Multiple DNA Targets

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• 作者：Jinchao Guo ; Litao Yang ; Lili Chen ; Dany Morisset ; Xiang Li ; Liangwen Pan ; Dabing Zhang
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：March 1, 2011
• 年：2011
• 卷：83
• 期：5
• 页码：1579-1586
• 全文大小：1047K
• 年卷期：v.83,no.5(March 1, 2011)
• ISSN：1520-6882

文摘

We describe the development of a novel combined approach for high-throughput analysis of multiple DNA targets based on multiplex Microdroplet PCR Implemented Capillary gel electrophoresis (MPIC), a two-step PCR amplification strategy. In the first step, the multiple target DNAs are preamplified using bipartite primers attached with universal tail sequences on their 5鈥?ends. Then, the preamplified templates are compartmentalized individually in the microdroplet of the PCR system, and multiple targets can be amplified in parallel, employing primers targeting their universal sequences. Subsequently, the resulting multiple products are analyzed by capillary gel electrophoresis (CGE). Using genetically modified organism (GMO) analysis as a model, 24 DNA targets can be simultaneously detected with a relative limit of detection of 0.1% (w/w) and absolute limit of detection of 39 target DNA copies. The described system provides a promising alternative for high-throughput analysis of multiple DNA targets.