title: Transcriptome analysis device based on liquid phase detection by fluorescently labeled nucleic acid probes.
abstract: 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.
authors.affiliation: Department of Microsystem Technology, Ritsumeikan University, 1-1-1 Noji-higashi, Kusatsu, Shiga, Japan. ryuji@se.ritsumei.ac.jp
authors.forename: Ryuji
authors.initials: R
authors.lastname: Yokokawa
authors.forename: Soichiro
authors.initials: S
authors.lastname: Tamaoki
authors.forename: Takashi
authors.initials: T
authors.lastname: Sakamoto
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
authors.forename: Susumu
authors.initials: S
authors.lastname: Sugiyama
mesh_terms: D004867:Equipment Design; D019544:Equipment Failure Analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D044367:Phase Transition; D015347:RNA Probes / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods; D014157:Transcription Factors / Q000032:analysis* / Q000235:genetics*
chemical_list: D015347:RNA Probes; D012333:RNA, Messenger; D014157:Transcription Factors