title: Integrated allele-specific polymerase chain reaction-capillary electrophoresis microdevice for single nucleotide polymorphism genotyping.
abstract: An integrated allele-specific (AS) polymerase chain reaction (PCR) and capillary electrophoresis (CE) microdevice has been developed for multiplex single nucleotide polymorphism (SNP) genotyping on a portable instrumentation, which was applied for on-site identification of HANWOO (Korean indigenous beef cattle). Twelve sets of primers were designed for targeting beef cattle's eleven SNP loci for HANWOO verification and one primer set for a positive PCR control, and the success rate for identification of HANWOO was demonstrated statistically. The AS PCR and CE separation for multiplex SNP typing was carried out on a glass-based microchip consisting of four layers: a microchannel plate for microfluidic control, a Pt-electrode plate for a resistance temperature detector (RTD), a poly(dimethylsiloxane) (PDMS) membrane and a manifold glass for microvalve function. The operation of the sample loading, AS PCR, microvalve, and CE on a chip was automated with a portable genetic analyzer, and the laser-induced fluorescence detection was performed on a miniaturized fluorescence detector. The blind samples were correctly identified as a HANWOO by showing one or two amplicon peaks in the electropherogram, while the imported beef cattle revealed more than five peaks. Our genetic analysis platform provides rapid, accurate, and on-site multiplex SNP typing.
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Program) and KAIST Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jong Young
authors.initials: JY
authors.lastname: Choi
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Program) and KAIST Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Yong Tae
authors.initials: YT
authors.lastname: Kim
authors.affiliation: Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Dahak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jinwoo
authors.initials: J
authors.lastname: Ahn
authors.affiliation: College of Agriculture, Life and Environment Sciences, Chungbuk National University, 52 Naesudong-ro, Heungdeok-gu, Cheongju, Chungbuk 361-763, Republic of Korea.
authors.forename: Kwan Suk
authors.initials: KS
authors.lastname: Kim
authors.affiliation: Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Dahak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Dae-Gab
authors.initials: DG
authors.lastname: Gweon
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Program) and KAIST Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: seots@kaist.ac.kr.
authors.forename: Tae Seok
authors.initials: TS
authors.lastname: Seo
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000145:classification / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D005787:Gene Frequency; D060005:Genotyping Techniques / Q000295:instrumentation* / Q000379:methods; D056656:Lab-On-A-Chip Devices; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D056910:Republic of Korea; D013045:Species Specificity
chemical_list: D017931:DNA Primers