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 language | English |
|---|---|
| Pages (from-to) | 1878-1882 |
| Number of pages | 5 |
| Journal | Biosensors and Bioelectronics |
| Volume | 23 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 2008 Jul 15 |
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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 journal › Article
}
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 -