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Bioinformatics Advance Access published online on May 30, 2007
Bioinformatics, doi:10.1093/bioinformatics/btm271
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© The Author (2007). Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Short oligonucleotide probes containing G-stacks display abnormal binding affinity on Affymetrix microarrays

Chunlei Wu 1,2,3, Haitao Zhao 2, Keith Baggerly 2,3, Roberto Carta 4 and Li Zhang 2,3,*

1 Genomic Institute of Novartis Research Foundation, 10675 John Jay Hopkins Dr, San Diego, CA 92121, USA. 2Department of Bioinformatics and Computational Biology, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Box 237, Houston, TX 77030, USA. 3Program in Biomathematics & Biostatistics, The University of Texas Graduate School of Biomedical Sciences at Houston, 6767 Bertner Avenue, Houston, TX 77225-0334, USA. 4Department of Statistics and Actuarial Sciences, University of Central Florida, Orlando, FL 32816, USA.

*To whom correspondence should be addressed. Dr. Li Zhang, E-mail: lzhangli{at}mdanderson.org


  Abstract

Motivation: In microarray experiments, probe design is critical to the specific and accurate measurement of target concentrations. Current designs select suitable probes through in silico scanning of transcriptome/genome based on first principles. However, due to lack of tools, the observed microarray data have not been used to assess the performance of individual probes to provide feedback to improve future designs.

Results: In this study, we describe a probe performance assessment method based on the concordance of the observed signals from probes that share common targets. Using this method, we found that probes containing multiple guanines in a row (G-stacks) have abnormal binding behavior compared with other probes, both in gene expression assays and genotyping assays using Affymetrix microarrays. These probes are less likely to covary with other probes that interrogate the same genes. Moreover, we found that these probes are much more likely to produce outliers when fitting the observed signals according to the Positional Dependent Nearest-Neighbor model, which gives reasonable estimates of binding affinity for most other probes. These results suggest that probes containing G-stacks tend to have increased cross hybridization signals and reduced target-specific hybridization signals, presumably due to multiplex binding forming G-quartet structures. Our findings are expected to be useful in microarray design and data analysis.

Availability: URL: http://odin.mdacc.tmc.edu/~zhangli/PerfectMatch/ contains the computer program for calculating correlations of neighboring probes.

Supplementary Material: Bioinformatics Online or http://odin.mdacc.tmc.edu/~zhangli/G-stack

Associate Editor: Prof. Martin Bishop

Received on February 1, 2007; revised on April 22, 2007; accepted on May 11, 2007

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