title: Triblock copolymer-based microchip device for rapid analysis of stuffer-free multiplex ligation-dependent probe amplification products.
abstract: Recent improvements in the multiplex ligation-dependent probe amplification (MLPA) method promise successful multiplex analysis of various genetic markers. In particular, it has been demonstrated that elimination of the stuffer sequence included in MLPA probes for length-dependent analysis substantially simplifies the probe design process and improves the accuracy of the analysis. As is the case for other CE-based methods, MLPA could be further developed on a microchip platform. However, high-resolution analysis of short MLPA probes requires careful microchip operation. In this study, we developed a microchip device for the multiplex analysis of five food-borne pathogens using a stuffer-free probe set. Microchip channel design and electrophoresis operating conditions were first optimized for reproducible analysis, after which two sieving matrices were tested. Finally, the method was validated using DNA samples isolated from intentionally infected milk.
authors.affiliation: Institute of Environmental and Energy Technology, Pohang University of Science and Technology, Pohang, Gyeongbuk, Korea.
authors.forename: Gi Won
authors.initials: GW
authors.lastname: Shin
authors.forename: Yong Tae
authors.initials: YT
authors.lastname: Kim
authors.forename: Hyun Young
authors.initials: HY
authors.lastname: Heo
authors.forename: Boram
authors.initials: B
authors.lastname: Chung
authors.forename: Tae Seok
authors.initials: TS
authors.lastname: Seo
authors.forename: Gyoo Yeol
authors.initials: GY
authors.lastname: Jung
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D008892:Milk / Q000382:microbiology; D058889:Molecular Typing / Q000295:instrumentation / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry*; D018807:Polymorphism, Single-Stranded Conformational; D011409:Propylene Glycols / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; C116176:PEO-PPO-PEO; D011409:Propylene Glycols; D011092:Polyethylene Glycols