title: Self-self hybridization as an alternative experiment design to dye swap for two-color microarrays.
abstract: Dye-specific bias effects, commonly observed in the two-color microarray platform, are normally corrected using the dye swap design. This design, however, is relatively expensive and labor-intensive. We propose a self-self hybridization design as an alternative to the dye swap design. In this design, the treated and control samples are labeled with Cy5 and Cy3 (or Cy3 and Cy5), respectively, without dye swap, along with a set of self-self hybridizations on the control sample. We compare this design with the dye swap design through investigation of mouse primary hepatocytes treated with three peroxisome proliferator-activated receptor-alpha (PPARalpha) agonists at three dose levels. Using Agilent's Whole Mouse Genome microarray, differentially expressed genes (DEG) were determined for both the self-self hybridization and dye swap designs. The DEG concordance between the two designs was over 80% across each dose treatment and chemical. Furthermore, 90% of DEG-associated biological pathways were in common between the designs, indicating that biological interpretations would be consistent. The reduced labor and expense for the self-self hybridization design make it an efficient substitute for the dye swap design. For example, in larger toxicogenomic studies, only about half the chips are required for the self-self hybridization design compared to that needed in the dye swap design.
authors.affiliation: Division of Bioinformatics, Z-Tech Corporation, Jefferson, Arkansas, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Fang
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Fan
authors.forename: Lei
authors.initials: L
authors.lastname: Guo
authors.forename: Leming
authors.initials: L
authors.lastname: Shi
authors.forename: Roger
authors.initials: R
authors.lastname: Perkins
authors.forename: Weigong
authors.initials: W
authors.lastname: Ge
authors.forename: Yvonne P
authors.initials: YP
authors.lastname: Dragan
authors.forename: Weida
authors.initials: W
authors.lastname: Tong
mesh_terms: D000818:Animals; D002232:Carbocyanines / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D005786:Gene Expression Regulation; D016678:Genome; D023281:Genomics; D022781:Hepatocytes / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; C089187:cyanine dye 3; C085321:cyanine dye 5