韩牛
基因分型
放大器
多重聚合酶链反应
聚合酶链反应
多路复用
毛细管电泳
打字
单核苷酸多态性
分子生物学
生物
基因型
SNP基因分型
电泳图谱
底漆(化妆品)
遗传学
化学
基因
有机化学
食品科学
作者
Jong Young Choi,Yong Tae Kim,Jin-Woo Ahn,Kwan Suk Kim,Dae-Gab Gweon,Tae Seok Seo
标识
DOI:10.1016/j.bios.2012.03.009
摘要
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.
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