Transcriptome analysis device based on liquid phase detection by fluorescently labeled nucleic acid probes.
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.
Yokokawa
Ryuji
R
Department of Microsystem Technology, Ritsumeikan University, 1-1-1 Noji-higashi, Kusatsu, Shiga, Japan. ryuji@se.ritsumei.ac.jp
Tamaoki
Soichiro
S
Sakamoto
Takashi
T
Murakami
Akira
A
Sugiyama
Susumu
S
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*
D015347:RNA Probes; D012333:RNA, Messenger; D014157:Transcription Factors