Bioinformatics Vol. 18 no. 11 2002
Pages 1470-1476
© 2002 Oxford University Press
Theoretical and experimental comparisons of gene expression indexes for oligonucleotide arrays
Division of Human Cancer Genetics, The Ohio State University, Columbus, Ohio, USA
Received on September 18, 2001
; revised on April 22, 2002
; accepted on April 30, 2002
Motivation: Oligonucleotide expression arrays exhibit systematic and reproducible variation produced by the multiple distinct probes used to represent a gene. Recently, a gene expression index has been proposed that explicitly models probe effects, and provides improved fits of hybridization intensity for arrays containing perfect match (PM) and mismatch (MM) probe pairs.
Results: Here we use a combination of analytical arguments and empirical data to show directly that the estimates provided by model-based expression indexes are superior to those provided by commercial software. The improvement is greatest for genes in which probe effects vary substantially, and modeling the PM and MM intensities separately is superior to using the PM–MM differences. To empirically compare expression indexes, we designed a mixing experiment involving three groups of human fibroblast cells (serum starved, serum stimulated, and a 50:50 mixture of starved/stimulated), with six replicate HuGeneFL arrays in each group. Careful spiking of control genes provides evidence that 88–98% of the genes on the array are detectably transcribed, and that the model-based estimates can accurately detect the presence versus absence of a gene. The use of extensive replication from single RNA sources enables exploration of the technical variability of the array.
Availability: Scripts for computing the Li–Wong reduced and full models are available in C, Splus and Perl in the supplementary information.
Contact: fwright{at}bios.unc.edu
Supplementary information: http://thinker.med.ohio-state.edu
* To whom correspondence should be addressed at Present address: Department of Biostatistics UNC-CH, 3107B McGavran-Greenberg Hall, CB #7420, Chapel Hill, NC 27599-7420, USA
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