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PubMed · 15123585

A versatile assay for high-throughput gene expression profiling on universal array matrices.

Jian-Bing Fan·Joanne M Yeakley·Marina Bibikova·Eugene Chudin·Eliza Wickham·Jing Chen·Dennis Doucet·Philippe Rigault·Baohong Zhang·Richard Shen·Celeste McBride·Hai-Ri Li·Xiang-Dong Fu·Arnold Oliphant·David L Barker·Mark S Chee
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Abstract

We report a flexible, sensitive, and quantitative gene-expression profiling system for assaying more than 400 genes, with three probes per gene, for 96 samples in parallel. The cDNA-mediated annealing, selection, extension and ligation (DASL) assay targets specific transcripts, using oligonucleotides containing unique address sequences that can hybridize to universal arrays. Cell-specific gene expression profiles were obtained using this assay for hormone-treated cell lines and laser-capture microdissected cancer tissues. Gene expression profiles derived from this assay were consistent with those determined by qRT-PCR. The DASL assay has been automated for use with a bead-based 96-array matrix system. The combined high-throughput assay and readout system is accurate and efficient, and can cost-effectively profile the expression of hundreds of genes in thousands of samples.

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AndrogensAnimalsCell LineCell Line, TumorDNA, ComplementaryGene Expression ProfilingGene Expression RegulationHumansLasersMaleMiceMicrodissectionOligonucleotide Array Sequence AnalysisOrgan SpecificityProstatic NeoplasmsRNAReverse Transcriptase Polymerase Chain ReactionSensitivity and SpecificityComparative StudyJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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Jian-Bing Fan, Joanne M Yeakley, Marina Bibikova, Eugene Chudin, Eliza Wickham, Jing Chen, Dennis Doucet, Philippe Rigault, Baohong Zhang, Richard Shen, Celeste McBride, Hai-Ri Li, Xiang-Dong Fu, Arnold Oliphant, David L Barker, Mark S Chee. 2004. A versatile assay for high-throughput gene expression profiling on universal array matrices.. https://doi.org/10.1101/gr.2167504

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