Fast and sensitive DNA analysis using changes in the FRET signals of molecular beacons in a PDMS microfluidic channel
- Authors
- Jung, Jaehyun; Chen, Lingxin; Lee, Sangyup; Kim, Sungyong; Seong, Gi Hun; Choo, Jaebum; Lee, Eun Kyu; Oh, Chil-Hwan; Lee, Sanghoon
- Issue Date
- Apr-2007
- Publisher
- SPRINGER HEIDELBERG
- Keywords
- fluorescence resonance energy transfer; molecular beacon; lab-on-a-chip; surface-enhanced Raman; DNA hybridization
- Citation
- ANALYTICAL AND BIOANALYTICAL CHEMISTRY, v.387, no.8, pp 2609 - 2615
- Pages
- 7
- Indexed
- SCIE
SCOPUS
- Journal Title
- ANALYTICAL AND BIOANALYTICAL CHEMISTRY
- Volume
- 387
- Number
- 8
- Start Page
- 2609
- End Page
- 2615
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/43810
- DOI
- 10.1007/s00216-007-1158-6
- ISSN
- 1618-2642
1618-2650
- Abstract
- A new DNA hybridization analytical method using a microfluidic channel and a molecular beacon-based probe (MB-probe) is described. A stem-loop DNA oligonucleotide labeled with two fluorophores at the 5' and 3' termini (a donor dye, TET, and an acceptor dye, TAMRA, respectively) was used to carry out a fast and sensitive DNA analysis. The MB-probe utilized the specificity and selectivity of the DNA hairpin-type probe DNA to detect a specific target DNA of interest. The quenching of the fluorescence resonance energy transfer (FRET) signal between the two fluorophores, caused by the sequence-specific hybridization of the MB-probe and the target DNA, was used to detect a DNA hybridization reaction in a poly(dimethylsiloxane) (PDMS) microfluidic channel. The azoospermia gene, DYS 209, was used as the target DNA to demonstrate the applicability of the method. A simple syringe pumping system was used for quick and accurate analysis. The laminar flow along the channel could be easily controlled by the 3-D channel structure and flow speed. By injecting the MB-probe and target DNA solutions into a zigzag-shaped PDMS microfluidic channel, it was possible to detect their sequence-specific hybridization. Surface-enhanced Raman spectroscopy (SERS) was also used to provide complementary evidence of the DNA hybridization. Our data show that this technique is a promising real-time detection method for label-free DNA targets in the solution phase.
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