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

@article{citeulike:1902246, 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. Result: 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. Contact: lzhangli@mdanderson.org Supplementary information: Bioinformatics online or http://odin.mdacc.tmc.edu/\~{}zhangli/G-stack 10.1093/bioinformatics/btm271}, author = {Wu, Chunlei and Zhao, Haitao and Baggerly, Keith and Carta, Roberto and Zhang, Li }, citeulike-article-id = {1902246}, doi = {http://dx.doi.org/10.1093/bioinformatics/btm271}, journal = {Bioinformatics}, keywords = {affy, low\_level}, month = {October}, number = {19}, pages = {2566--2572}, posted-at = {2008-02-04 11:50:29}, priority = {1}, title = {Short oligonucleotide probes containing G-stacks display abnormal binding affinity on Affymetrix microarrays}, url = {http://dx.doi.org/10.1093/bioinformatics/btm271}, volume = {23}, year = {2007} }

TY - JOUR ID - citeulike:1902246 L3 - citeulike-article-id:1902246 TI - Short oligonucleotide probes containing G-stacks display abnormal binding affinity on Affymetrix microarrays JF - Bioinformatics VL - 23 IS - 19 SP - 2566 EP - 2572 N2 - 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. Result: 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. Contact: lzhangli@mdanderson.org Supplementary information: Bioinformatics online or http://odin.mdacc.tmc.edu/~zhangli/G-stack 10.1093/bioinformatics/btm271 KW - affy KW - low_level AU - Wu, Chunlei AU - Zhao, Haitao AU - Baggerly, Keith AU - Carta, Roberto AU - Zhang, Li PY - 2007/10/01/ UR - http://dx.doi.org/10.1093/bioinformatics/btm271 ER -