Transcriptome analysis device based on liquid phase detection by fluorescently labeled nucleic acid probes
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Personalized medicine based on genetic information has been proposed as an attractive medical treatment. It is supported by the rapid development of the gene diagnosis utilizing on-chip analysis technology. This study reports characterizations of mRNA detection device by its hybridization with 2′-O-methyl oligoribonucleotide probe in a liquid phase, which eliminates time-consuming processes such as removing non-hybridized probes in conventional solid phase detection. In order to achieve a high sensitivity in the fluorescent detection of the target mRNA, we have optimized detection channel depth and width for a microfluidic device, and investigated polydimethyl siloxiane components and observation setup to minimize background noise. Adopting optimized channel dimensions and setup for the probe, the fluorescent intensity increased 3.3-fold at the lowest detectable concentration of 7.8 nM.
KeywordsMicroTAS Fluorescent detection Microfluidic device Hybridization mRNA
PDMS components were supplied by Shin-Etsu Chemical Co., Ltd. This work was carried out under the 21st century COE program “Micro/Nano Science and Integrated Systems” supported by the Ministry of Education and Kyoto nanotechnology cluster supported by the Advanced Software Technology and Mechatronics Research Institute.
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