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Proceedings Paper

Label-free DNA hybridization detection with molecular beacon immobilized in photopolymerized acrylamide gel microarray
Author(s): Hong Wang; Jiong Li; Quanjun Liu; Heping Liu; Zuhong Lu; Jijun Zhu
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Paper Abstract

Photopolymerized acrylamide gel microarray used to immobilize the molecular beacon for label free DNA hybridization detection is described in this paper. Polyacrylamide gel microarray was prepared by UV photopolymerization of 4% acrylamide in 40% glycerol, 0.002%methylene blue, 0.012% TEMED and 0.1M phosphate buffer (PHequals7) with a Relpel-silane pretreated quartz mask. This kind of three-dimensional gel microarray provides more than 100 times great immobilization capacity. The hybridization and other processes with it resemble a homogeneous liquid phase reaction rather than a heterogeneous liquid-solid interface reaction. The specially designed molecular beacons contain a 15 base loop sequence with 5 base pair stem, a 20 base thymine as spacer, a 5'-end amino group for immobilization, a fluorescein in the middle of the sequence as the fluorophore and a 3'-end DABCYL as the quencher. Between the 5'-end amino group and the stem, the 20 base thymine is used to minimize destability caused by 5'-end immobilization. Confocal microscope was used to investigate the fluorescence intensity of gel immobilized molecular beacon probes. After hybridization we can easily distinguish complementary and noncomplementay targets with gel-immobilized molecular beacon probes. Image analysis showed that the fluorescence intensity ratio of complementary to noncomplementay probes is great than 5. The potential applications of gel-immobilized molecular beacon microarray are mutation detection, pathogenic detection, etc. in a parallel, cost saving and label-free detection way.

Paper Details

Date Published: 15 October 2001
PDF: 4 pages
Proc. SPIE 4601, Micromachining and Microfabrication Process Technology and Devices, (15 October 2001); doi: 10.1117/12.444684
Show Author Affiliations
Hong Wang, Southeast Univ. (China)
Jiong Li, Southeast Univ. (China)
Quanjun Liu, Southeast Univ. (China)
Heping Liu, Southeast Univ. (China)
Zuhong Lu, Southeast Univ. (China)
Jijun Zhu, Southeast Univ. (China)

Published in SPIE Proceedings Vol. 4601:
Micromachining and Microfabrication Process Technology and Devices
Norman C. Tien; Qing-An Huang, Editor(s)

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