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Nucleic Acids Research 2004 32(18):e137; doi:10.1093/nar/gnh133
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Published online 8 October 2004

Nucleic Acids Research, Vol. 32 No. 18 © Oxford University Press 2004; all rights reserved

Gold nanoparticle probe-based gene expression analysis with unamplified total human RNA

Martin Huber, Tai-Fen Wei, Uwe R. Müller, Phil A. Lefebvre, Sudhakar S. Marla and Y. Paul Bao*

Nanosphere Inc., 4088 Commercial Ave., Northbrook, IL 60062, USA

* To whom correspondence should be addressed. Tel: +1 847 400 9150; Fax: +1 847 400 9199; Email: pbao{at}nanosphere.us

Received July 6, 2004; Revised and Accepted September 13, 2004

Microarray-based gene expression analysis plays a pivotal role in modern biology and is poised to enter the field of molecular diagnostics. Current microarray-based gene expression systems typically require enzymatic conversion of mRNA into labeled cDNA or cRNA. Conversion to cRNA involves a target amplification step that overcomes the low sensitivity associated with commonly used fluorescent detection methods. Herein, we present a novel enzyme-free, microarray-based gene expression system that uses unamplified total human RNA sample as the target nucleic acid. The detection of microarray-bound RNA molecules is accomplished by targeting the poly-A tail with an oligo-dT20 modified gold nanoparticle probe, signal amplification by autometallography, and subsequent measurement of nanoparticle-mediated light scattering. The high sensitivity afforded by the nanoparticle probes allows differential gene expression from as little as 0.5 µg unamplified total human RNA in a 2 h hybridization without the need for elaborate sample labeling steps.


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