title: MPIC: a high-throughput analytical method for multiple DNA targets.
abstract: 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'-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.
authors.affiliation: GMO Detection Laboratory, SJTU-Bor Luh Food Safety Center, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
authors.forename: Jinchao
authors.initials: J
authors.lastname: Guo
authors.forename: Litao
authors.initials: L
authors.lastname: Yang
authors.forename: Lili
authors.initials: L
authors.lastname: Chen
authors.forename: Dany
authors.initials: D
authors.lastname: Morisset
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Liangwen
authors.initials: L
authors.lastname: Pan
authors.forename: Dabing
authors.initials: D
authors.lastname: Zhang
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D017931:DNA Primers; D019075:Electrophoresis, Capillary / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D004247:DNA