DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes

Lingxin Chen, Sangyeop Lee, Moonkwon Lee, Chaesung Lim, Jaebum Choo, Joong Yull Park, Sanghoon Lee, Sang Woo Joo, Kyeong Hee Lee, Young Wook Choi

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

A conceptually new technique for fast DNA detection has been developed. Here, we report a fast and sensitive online fluorescence resonance energy transfer (FRET) detection technique for label-free target DNA. This method is based on changes in the FRET signal resulting from the sequence-specific hybridization between two fluorescently labelled nucleic acid probes and target DNA in a PDMS microfluidic channel. Confocal laser-induced microscopy has been used for the detection of fluorescence signal changes. In the present study, DNA hybridizations could be detected without PCR amplification because the sensitivity of confocal laser-induced fluorescence detection is very high. Two probe DNA oligomers (5′-CTGAT TAGAG AGAGAA-TAMRA-3′ and 5′-TET-ATGTC TGAGC TGCAGG-3′) and target DNA (3′-GACTA ATCTC TCTCT TACAG GCACT ACAGA CTCGA CGTCC-5′) were introduced into the channel by a microsyringe pump, and they were efficiently mixed by passing through the alligator teeth-shaped PDMS microfluidic channel. Here, the nucleic acid probes were terminally labelled with the fluorescent dyes, tetrafluororescein (TET) and tetramethyl-6-carboxyrhodamine (TAMRA), respectively. According to our confocal fluorescence measurements, the limit of detection of the target DNA is estimated to be 1.0 × 10-6 to 1.0 × 10-7 M. Our result demonstrates that this analytical technique is a promising diagnostic tool that can be applied to the real-time analysis of DNA targets in the solution phase.

Original languageEnglish
Pages (from-to)1878-1882
Number of pages5
JournalBiosensors and Bioelectronics
Volume23
Issue number12
DOIs
Publication statusPublished - 2008 Jul 15

Keywords

  • Confocal laser-scanning microscopy
  • DNA hybridization
  • FRET
  • Microfluidic sensor
  • Real-time analysis

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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  • Cite this

    Chen, L., Lee, S., Lee, M., Lim, C., Choo, J., Park, J. Y., Lee, S., Joo, S. W., Lee, K. H., & Choi, Y. W. (2008). DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes. Biosensors and Bioelectronics, 23(12), 1878-1882. https://doi.org/10.1016/j.bios.2008.02.013