Independence and reproducibility across microarray platforms.
Microarrays have been widely used for the analysis of gene expression, but the issue of reproducibility across platforms has yet to be fully resolved. To address this apparent problem, we compared gene expression between two microarray platforms: the short oligonucleotide Affymetrix Mouse Genome 430 2.0 GeneChip and a spotted cDNA array using a mouse model of angiotensin II-induced hypertension. RNA extracted from treated mice was analyzed using Affymetrix and cDNA platforms and then by quantitative RT-PCR (qRT-PCR) for validation of specific genes. For the 11,710 genes present on both arrays, we assessed the relative impact of experimental treatment and platform on measured expression and found that biological treatment had a far greater impact on measured expression than did platform for more than 90% of genes, a result validated by qRT-PCR. In the small number of cases in which platforms yielded discrepant results, qRT-PCR generally did not confirm either set of data, suggesting that sequence-specific effects may make expression predictions difficult to make using any technique.
Larkin
Jennie E
JE
Institute for Genomic Research, 9712 Medical Center Drive, Rockville, Maryland 20850, USA.
Frank
Bryan C
BC
Gavras
Haralambos
H
Sultana
Razvan
R
Quackenbush
John
J
D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D020869:Gene Expression Profiling / Q000592:standards; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards*; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017395:Plasminogen Activator Inhibitor 1; D000804:Angiotensin II