Automation of cDNA microarray hybridization and washing yields improved data quality.
Microarray technology allows the analysis of whole-genome transcription within a single hybridization, and has become a standard research tool. It is extremely important to minimize variation in order to obtain high quality microarray data that can be compared among experiments and laboratories. The majority of facilities implement manual hybridization approaches for microarray studies. We developed an automated method for cDNA microarray hybridization that uses equivalent pre-hybridization, hybridization and washing conditions to the suggested manual protocol. The automated method significantly decreased variability across microarray slides compared to manual hybridization. Although normalized signal intensities for buffer-only spots across the chips were identical, significantly reduced variation and inter-quartile ranges were obtained using the automated workstation. This decreased variation led to improved correlation among technical replicates across slides in both the Cy3 and Cy5 channels.
Yauk
Carole
C
Environmental and Occupational Toxicology Division, Health Canada, Ottawa, Ontario, Canada. carole_yauk@hc-sc.gc.ca
Berndt
Lynn
L
Williams
Andrew
A
Douglas
George R
GR
D000818:Animals; D002232:Carbocyanines / Q000737:chemistry; D002460:Cell Line; D020869:Gene Expression Profiling / Q000379:methods; D051379:Mice; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D015203:Reproducibility of Results
D002232:Carbocyanines; C089187:cyanine dye 3; C085321:cyanine dye 5