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Proceedings Paper

Mosaic DNA chip fabrication and its time-resolved fluorescence detection
Author(s): Quanguo He; Hong Chen; Jianxin Tang; Pengfeng Xiao; Nongyue He
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Paper Abstract

Lab-on-a-Chip (LOC) and μ-TAS (micro-total analytical system) are based on miniaturized integrated platforms that have the potential to revolutionize chemical, biological, and biochemical synthesis and analysis. Here, we demonstrated a process of fabricating a mosaic DNA chip and a corresponding detection method by time-resolved fluorescence (TRF) labeling. We synthesized oligonucleotide sequences in situ on glass slides directly, and then sliced them up into small pieces and patched up the pieces with different sequences to generate a mosaic DNA chip. With multiple BCPDA (BCPDA, abbreviated from 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid) labeling method based on biotin-avidin amplification, we established a TRF detection format on the mosaic DNA chip. The detection method allows discriminatory signals for perfect match, one-base mismatch, two-base mismatch and three-base mismatch by TRF labeled hybridization, whereby Europium (III, Eu3+) was captured and released on the principle of complexation and dissociation interaction between BCPDA and Eu3+ solution when the BCPDA-tagged avidin and biotin-ended oligonucleotide sequence linked. The fluorescence spectra and related lifetimes were determined. Also, we compared the TRF detection mode with the conventional fluorescence one. These results showed the former is more reliable and stable than the latter, especially for the mosaic DNA chip. Likewise, by applying TRF probing (or labeling) to specific bio-systems, the discovery is of fundamental interest and has significant implications to time-resolved-fluorescence based detection on biosensor.

Paper Details

Date Published: 29 December 2004
PDF: 10 pages
Proc. SPIE 5593, Nanosensing: Materials and Devices, (29 December 2004); doi: 10.1117/12.569578
Show Author Affiliations
Quanguo He, Zhuzhou Institute of Technology (China)
Southeast Univ. (China)
Central South Forestry Univ. (China)
Hong Chen, Zhuzhou Institute of Technology (China)
Central South Forestry Univ. (China)
Jianxin Tang, Zhuzhou Institute of Technology (China)
Pengfeng Xiao, Southeast Univ. (China)
Nongyue He, Zhuzhou Institute of Technology (China)

Published in SPIE Proceedings Vol. 5593:
Nanosensing: Materials and Devices
M. Saif Islam; Achyut K. Dutta, Editor(s)

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