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, Sang Hoon Lee, Sang Woo Joo, Kyeong Hee Lee, Young Wook Choi

Research output: Contribution to journalArticle

49 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

Fingerprint

Nucleic Acid Probes
Microfluidics
Nucleic acids
DNA
Fluorescence Resonance Energy Transfer
Fluorescence
Confocal Microscopy
Alligators and Crocodiles
Lasers
DNA Probes
Protein Sorting Signals
Fluorescent Dyes
Oligomers
Amplification
Limit of Detection
Labels
Tooth
Microscopic examination
Pumps
Dyes

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Electrochemistry

Cite this

DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes. / Chen, Lingxin; Lee, Sangyeop; Lee, Moonkwon; Lim, Chaesung; Choo, Jaebum; Park, Joong Yull; Lee, Sang Hoon; Joo, Sang Woo; Lee, Kyeong Hee; Choi, Young Wook.

In: Biosensors and Bioelectronics, Vol. 23, No. 12, 15.07.2008, p. 1878-1882.

Research output: Contribution to journalArticle

Chen, L, Lee, S, Lee, M, Lim, C, Choo, J, Park, JY, Lee, SH, Joo, SW, Lee, KH & Choi, YW 2008, 'DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes', Biosensors and Bioelectronics, vol. 23, no. 12, pp. 1878-1882. https://doi.org/10.1016/j.bios.2008.02.013
Chen, Lingxin ; Lee, Sangyeop ; Lee, Moonkwon ; Lim, Chaesung ; Choo, Jaebum ; Park, Joong Yull ; Lee, Sang Hoon ; Joo, Sang Woo ; Lee, Kyeong Hee ; Choi, Young Wook. / DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes. In: Biosensors and Bioelectronics. 2008 ; Vol. 23, No. 12. pp. 1878-1882.
@article{e0e73009e51c41cf937c267cae27187a,
title = "DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes",
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.",
keywords = "Confocal laser-scanning microscopy, DNA hybridization, FRET, Microfluidic sensor, Real-time analysis",
author = "Lingxin Chen and Sangyeop Lee and Moonkwon Lee and Chaesung Lim and Jaebum Choo and Park, {Joong Yull} and Lee, {Sang Hoon} and Joo, {Sang Woo} and Lee, {Kyeong Hee} and Choi, {Young Wook}",
year = "2008",
month = "7",
day = "15",
doi = "10.1016/j.bios.2008.02.013",
language = "English",
volume = "23",
pages = "1878--1882",
journal = "Biosensors",
issn = "0956-5663",
publisher = "Elsevier Limited",
number = "12",

}

TY - JOUR

T1 - DNA hybridization detection in a microfluidic channel using two fluorescently labelled nucleic acid probes

AU - Chen, Lingxin

AU - Lee, Sangyeop

AU - Lee, Moonkwon

AU - Lim, Chaesung

AU - Choo, Jaebum

AU - Park, Joong Yull

AU - Lee, Sang Hoon

AU - Joo, Sang Woo

AU - Lee, Kyeong Hee

AU - Choi, Young Wook

PY - 2008/7/15

Y1 - 2008/7/15

N2 - 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.

AB - 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.

KW - Confocal laser-scanning microscopy

KW - DNA hybridization

KW - FRET

KW - Microfluidic sensor

KW - Real-time analysis

UR - http://www.scopus.com/inward/record.url?scp=43449114777&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=43449114777&partnerID=8YFLogxK

U2 - 10.1016/j.bios.2008.02.013

DO - 10.1016/j.bios.2008.02.013

M3 - Article

C2 - 18378133

AN - SCOPUS:43449114777

VL - 23

SP - 1878

EP - 1882

JO - Biosensors

JF - Biosensors

SN - 0956-5663

IS - 12

ER -