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A qualitative assessment of direct-labeled cDNA products prior to microarray analysis.

Grissom SF, Lobenhofer EK, Tucker CJ

National Institute of Environmental Health Sciences Intramural Microarray Group, Research Triangle Park, NC 27709, USA. grissom2@niehs.nih.gov

BACKGROUND: The success of the microarray process in determining differential gene expression of thousands of genes is dependent upon the quality and integrity of the starting RNA, this being particularly true of direct labeling via a reverse transcription procedure. Furthermore, an RNA of reasonable quality still may not yield reliable hybridization data if the labeling efficiency was poor. RESULTS: Here we present a novel assay for assessing the quality of directly labeled fluorescent cDNA prior to microarray hybridization utilizing the Agilent 2100 Bioanalyzer, which employs microfluidic technology for the analysis of nucleic acids and proteins. Using varying amounts of RNase to simulate RNA degradation, we show the strength of this un-advertised assay in determining the relative amounts of cDNA obtained from a direct labeling reaction. CONCLUSION: Utilization of this method in the lab will help to prevent the costly mistake of hybridizing poor quality direct labeled products to expensive arrays.

Published 7 April 2005 in BMC Genomics, 6(1): 36.
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