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Limma: linear models for microarray data
- Bioinformatics and Computational Biology Solutions using R and Bioconductor
, 2005
"... This free open-source software implements academic research by the authors and co-workers. If you use it, please support the project by citing the appropriate journal articles listed in Section 2.1.Contents ..."
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Cited by 759 (13 self)
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This free open-source software implements academic research by the authors and co-workers. If you use it, please support the project by citing the appropriate journal articles listed in Section 2.1.Contents
Summaries of Affymetrix GeneChip probe level data
- Nucleic Acids Res
, 2003
"... High density oligonucleotide array technology is widely used in many areas of biomedical research for quantitative and highly parallel measurements of gene expression. Affymetrix GeneChip arrays are the most popular. In this technology each gene is typically represented by a set of 11±20 pairs of pr ..."
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Cited by 464 (21 self)
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High density oligonucleotide array technology is widely used in many areas of biomedical research for quantitative and highly parallel measurements of gene expression. Affymetrix GeneChip arrays are the most popular. In this technology each gene is typically represented by a set of 11±20 pairs of probes. In order to obtain expression measures it is necessary to summarize the probe level data. Using two extensive spike-in studies and a dilution study, we developed a set of tools for assessing the effectiveness of expression measures. We found that the performance of the current version of the default expression measure provided by Affymetrix Microarray Suite can be signi®cantly improved by the use of probe level summaries derived from empirically motivated statistical models. In particular, improvements in the ability to detect differentially expressed genes are demonstrated.
Normalization of cDNA microarray data
- Methods
, 2003
"... Normalization means to adjust microarray data for effects which arise from variation in the technology rather than from biological differences between the RNA samples or between the printed probes. This article describes normalization methods based on the fact that dye balance typically varies with ..."
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Cited by 241 (8 self)
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Normalization means to adjust microarray data for effects which arise from variation in the technology rather than from biological differences between the RNA samples or between the printed probes. This article describes normalization methods based on the fact that dye balance typically varies with spot intensity and with spatial position on the array. Print-tip loess normalization provides a well-tested general purpose normalization method which has given good results on a wide range of arrays. The method may be refined by using quality weights for individual spots. The method is best combined with diagnostic plots of the data which display the spatial and intensity trends. When diagnostic plots show that biases still remain in the data after normalization, further normalization steps such as plate-order normalization or scalenormalization between the arrays may be undertaken. Composite normalization may be used when control spots are available which are known to be not differentially expressed. Variations on loess normalization include global loess normalization and 2D normalization. Detailed commands are given to implement the normalization techniques using freely available software. 1
Quantitative quality control in microarray image processing and data acquisition
- Nucleic Acids Res
, 2001
"... experiments and the application in data filtering, normalization and false positive rate prediction ..."
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Cited by 115 (10 self)
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experiments and the application in data filtering, normalization and false positive rate prediction
Capturing heterogeneity in gene expression studies by ‘surrogate variable analysis’. PLoS Genetics 3:e161
, 2007
"... It has unambiguously been shown that genetic, environmental, demographic, and technical factors may have substantial effects on gene expression levels. In addition to the measured variable(s) of interest, there will tend to be sources of signal due to factors that are unknown, unmeasured, or too com ..."
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Cited by 108 (16 self)
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It has unambiguously been shown that genetic, environmental, demographic, and technical factors may have substantial effects on gene expression levels. In addition to the measured variable(s) of interest, there will tend to be sources of signal due to factors that are unknown, unmeasured, or too complicated to capture through simple models. We show that failing to incorporate these sources of heterogeneity into an analysis can have widespread and detrimental effects on the study. Not only can this reduce power or induce unwanted dependence across genes, but it can also introduce sources of spurious signal to many genes. This phenomenon is true even for well-designed, randomized studies. We introduce ‘‘surrogate variable analysis’ ’ (SVA) to overcome the problems caused by heterogeneity in expression studies. SVA can be applied in conjunction with standard analysis techniques to accurately capture the relationship between expression and any modeled variables of interest. We apply SVA to disease class, time course, and genetics of gene expression studies. We show that SVA increases the biological accuracy and reproducibility of analyses in genome-wide expression studies.
CLICK and EXPANDER: a system for clustering and visualizing gene expression data
- Bioinformatics
, 2003
"... Motivation: Microarrays have become a central tool in biological research. Their applications range from functional annotation to tissue classification and genetic network inference. A key step in the analysis of gene expression data is the identification of groups of genes that manifest similar exp ..."
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Cited by 101 (6 self)
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Motivation: Microarrays have become a central tool in biological research. Their applications range from functional annotation to tissue classification and genetic network inference. A key step in the analysis of gene expression data is the identification of groups of genes that manifest similar expression patterns. This translates to the algorithmic problem of clustering genes based on their expression patterns. Results: We present a novel clustering algorithm, called CLICK, and its applications to gene expression analysis. The algorithm utilizes graph-theoretic and statistical techniques to identify tight groups (kernels) of highly similar elements, which are likely to belong to the same true cluster. Several heuristic procedures are then used to expand the kernels into the full clusters. We report on the application of CLICK to a variety of gene expression data sets. In all those applications it outperformed extant algorithms according to several common figures of merit. We also point out that CLICK can be successfully used for the identification of common regulatory motifs in the upstream regions of co-regulated genes. Furthermore, we demonstrate how CLICK can be used to accurately classify tissue samples into disease types, based on their expression profiles. Finally, we present a new java-based graphical tool, called EXPANDER, for gene expression analysis and visualization, which incorporates CLICK and several other popular clustering algorithms.
lincRNAs act in the circuitry controlling pluripotency and differentiation. Nature 2011
"... and differentiation ..."
Statistical Issues in cDNA Microarray Data Analysis
, 2003
"... This article summarizes some of the issues involved and provides a brief review of the analysis tools which are available to researchers to deal with them. Any microarray experiment involves a number of distinct stages. Firstly there is the design of the experiment. The researchers must decide which ..."
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Cited by 84 (6 self)
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This article summarizes some of the issues involved and provides a brief review of the analysis tools which are available to researchers to deal with them. Any microarray experiment involves a number of distinct stages. Firstly there is the design of the experiment. The researchers must decide which genes are to be printed on the arrays, which sources of RNA are to be hybridized to the arrays and on how many arrays the hybridizations will be replicated. Secondly, after hybridization, there follows a number of data-cleaning steps or `low-level analysis' of the microarray data. The microarray images must be processed to acquire red and green foreground and background intensities for each spot. The acquired red/green ratios must be normalized to adjust for dye-bias and for any systematic variation other than that due to the differences between the RNA samples being studied. Thirdly, the normalized ratios are analyzed by various graphical and numerical means to select differentially expressed genes or to find groups of genes whose expression profiles can reliably classify the different RNA sources into meaningful groups. The sections of this article correspond roughly to the various analysis steps. The following notation will be used throughout the article. The foreground red and green intensities will be written Pp and 9p for each spot. The background intensities will be Pf and 9f . The background-corrected intensities will be P and 9 where usually P Pp Pf 0 # and 9 9p 9f 0 # . The log-differential expression ratio will be vyq # E P 9 0 for each spot. Finally, the log-intensity of the spot will be vyq 3 P9 0 , a measure of the overall brightness of the spot. (The letter E is a mnemonic for minus as vyq vyq E P 9 0 # while 3 is a mnemonic for add as #vyq vyq #...
Optimal Sample Size for Multiple Testing: the Case of Gene Expression Microarrays
- Journal of the American Statistical Association
, 2004
"... We consider the choice of an optimal sample size for multiple comparison problems. The motivating application is the choice of the number of microarray experiments to be carried out when learning about dierential gene expression. However, the approach is valid in any application that involves multip ..."
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Cited by 74 (5 self)
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We consider the choice of an optimal sample size for multiple comparison problems. The motivating application is the choice of the number of microarray experiments to be carried out when learning about dierential gene expression. However, the approach is valid in any application that involves multiple comparison in a large number of hypothesis tests.
Evaluation of gene expression measurements from commercial microarray platforms
- Nucleic Acids Res
, 2003
"... Multiple commercial microarrays for measuring genome-wide gene expression levels are currently available, including oligonucleotide and cDNA, single- and two-channel formats. This study reports on the results of gene expression measurements generated from identical RNA preparations that were obtaine ..."
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Cited by 71 (0 self)
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Multiple commercial microarrays for measuring genome-wide gene expression levels are currently available, including oligonucleotide and cDNA, single- and two-channel formats. This study reports on the results of gene expression measurements generated from identical RNA preparations that were obtained using three commercially available microarray platforms. RNA was collected from PANC-1 cells grown in serum-rich medium and at 24 h following the removal of serum. Three biological replicates were prepared for each condition, and three experimental replicates were produced for the ®rst biological replicate. RNA was labeled and hybridized to microarrays from three major suppliers according to manufacturers ' protocols, and gene expression measurements were obtained using each platform's standard software. For each platform, gene targets from a subset of 2009 common genes were compared. Correlations in gene expression levels and comparisons for signi®cant gene expression changes in this subset were calculated, and showed considerable divergence across the different platforms, suggesting the need for establishing industrial manufacturing standards, and further independent and thorough validation of the technology.
