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Fast and sensitive DNA analysis using changes in the FRET signals of molecular beacons in a PDMS microfluidic channel

Authors
Jung, JaehyunChen, LingxinLee, SangyupKim, SungyongSeong, Gi HunChoo, JaebumLee, Eun KyuOh, Chil-HwanLee, Sanghoon
Issue Date
4월-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
Indexed
SCIE
SCOPUS
Journal Title
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume
387
Number
8
Start Page
2609
End Page
2615
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/125791
DOI
10.1007/s00216-007-1158-6
ISSN
1618-2642
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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College of Medicine > Department of Medical Science > 1. Journal Articles
College of Health Sciences > School of Biomedical Engineering > 1. Journal Articles

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