title: Analysis of discordant Affymetrix probesets casts serious doubt on idea of microarray data reutilization.
abstract: BACKGROUND
  Affymetrix microarray technology allows one to investigate expression of thousands of genes simultaneously upon a variety of conditions. In a popular U133A microarray platform, the expression of 37% of genes is measured by more than one probeset. The discordant expression observed for two different probesets that match the same gene is a widespread phenomenon which is usually underestimated, ignored or disregarded.
  
  
  RESULTS
  Here we evaluate the prevalence of discordant expression in data collected using Affymetrix HG-U133A microarray platform. In U133A, about 30% of genes annotated by two different probesets demonstrate a substantial correlation between independently measured expression values. To our surprise, sorting the probesets according to the nature of the discrepancy in their expression levels allowed the classification of the respective genes according to their fundamental functional properties, including observed enrichment by tissue-specific transcripts and alternatively spliced variants. On another hand, an absence of discrepancies in probesets that simultaneously match several different genes allowed us to pinpoint non-expressed pseudogenes and gene groups with highly correlated expression patterns. Nevertheless, in many cases, the nature of discordant expression of two probesets that match the same transcript remains unexplained. It is possible that these probesets report differently regulated sets of transcripts, or, in best case scenario, two different sets of transcripts that represent the same gene.
  
  
  CONCLUSIONS
  The majority of absolute gene expression values collected using Affymetrix microarrays may not be suitable for typical interpretative downstream analysis.
authors.forename: Andrey
authors.initials: A
authors.lastname: Marakhonov
authors.forename: Nataliya
authors.initials: N
authors.lastname: Sadovskaya
authors.forename: Ivan
authors.initials: I
authors.lastname: Antonov
authors.forename: Ancha
authors.initials: A
authors.lastname: Baranova
authors.forename: Mikhail
authors.initials: M
authors.lastname: Skoblov
mesh_terms: D020869:Gene Expression Profiling; D006801:Humans; D058977:Molecular Sequence Annotation; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes
chemical_list: D015345:Oligonucleotide Probes