Gold nanoparticle probe-based gene expression analysis with unamplified total human RNA -: art. no. e137

被引:80
作者
Huber, M [1 ]
Wei, TF [1 ]
Müller, UR [1 ]
Lefebvre, PA [1 ]
Marla, SS [1 ]
Bao, YP [1 ]
机构
[1] Nanosphere Inc, Northbrook, IL 60062 USA
关键词
D O I
10.1093/nar/gnh133
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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-dT(20) 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 mug unamplified total human RNA in a 2 h hybridization without the need for elaborate sample labeling steps.
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页数:8
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