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

Abstract

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 microg unamplified total human RNA in a 2 h hybridization without the need for elaborate sample labeling steps.

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@article{Huber2004GoldNP, title={Gold nanoparticle probe-based gene expression analysis with unamplified total human RNA.}, author={M. C. E. Huber and T Wei and Uwe R. M{\"{u}ller and Phil A Lefebvre and Sudhakar S. Marla and Yijia P. Bao}, journal={Nucleic acids research}, year={2004}, volume={32 18}, pages={e137} }