Development of a microarray assay that measures hybridization stoichiometry in moles

Richard J.D. Rouse*, Celia R. Espinoza, R. Hannes Niedner, Gary Hardiman

*Corresponding author for this work

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Microarray data is most useful when it can be compared with other genetic detection technologies. In this report, we designed a microarray assay format that transforms raw data into a defined scientific unit (i.e., moles) by measuring the amount of array feature present and the cDNA sequence hybridized. This study profiles a mouse reference universal RNA sample on a microarray consisting of PCR products. In measuring array features, a labeled DNA sequence was designed that hybridizes to a conserved sequence that is present in every array feature. To measure the amount of cDNA sample hybridized, the RNA sample was processed to ensure consistent dye to DNA ratio for every labeled target cDNA molecule, using labeled branched dendrimers rather than by incorporation. A dye printing assay was then performed in order to correlate molecules of cyanine dye to signal intensity. We demonstrate that by using this microarray assay design, raw data can be transformed into defined scientific units, which will facilitate interpretation of other experiments, such as data deposited at the Gene Expression Omnibus and ArrayExpress.

Original languageEnglish
Pages (from-to)464-470
Number of pages7
JournalBioTechniques
Volume36
Issue number3
Publication statusPublished - 01 Mar 2004
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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  • Cite this

    Rouse, R. J. D., Espinoza, C. R., Niedner, R. H., & Hardiman, G. (2004). Development of a microarray assay that measures hybridization stoichiometry in moles. BioTechniques, 36(3), 464-470.