共查询到20条相似文献,搜索用时 15 毫秒
1.
Design and analysis of two-phase experiments for gene expression microarrays-part I. 总被引:1,自引:0,他引:1
Gene expression microarray experiments are intrinsically two-phase experiments. Messenger RNA (mRNA), required for the microarray experiment, must first be derived from plants or animals that are exposed to a set of treatments in a previous experiment (Phase 1). The mRNA is then used in the subsequent laboratory-based microarray experiment (Phase 2) from which gene expression is measured and ultimately analyzed. We show that obtaining a valid test for the effects of treatments on gene expression depends on the design of both the Phase 1 and Phase 2 experiments. Examples show that the multiple dye-swap design at Phase 2 is more robust than the alternating loop design in the absence of prior knowledge of the relative size of variation in the Phase 1 and Phase 2 experiments. 相似文献
2.
The major goal of two-color cDNA microarray experiments is to measure the relative gene expression level (i.e., relative amount of mRNA) of each gene between samples in studies of gene expression. More specifically, given an N-sample experiment, we need all N(N - 1)/2 relative expression levels of all sample pairs of each gene for identification of the differentially expressed genes and for clustering of gene expression patterns. However, the intensities observed from two-color cDNA microarray experiments do not simply represent the relative gene expression level. They are composed of signal (gene expression level), noise, and other factors. In discussions on the experimental design of two-color cDNA microarray experiments, little attention has been given to the fact that different combinations of test and control samples will produce microarray intensities data with varying intrinsic composition of factors. As a consequence, not all experimental designs for two-color cDNA microarray experiments are able to provide all possible relative gene expression levels. This phenomenon has never been addressed. To obtain all possible relative gene expression levels, a novel method for two-color cDNA microarray experimental design evaluation is necessary that will allow the making of an accurate choice. In this study, we propose a model-based approach to illustrate how the factor composition of microarray intensities changed with different experimental designs in two-color cDNA microarray experiments. By analyzing 12 experimental designs (including 5 general forms), we demonstrate that not all experimental designs are able to provide all possible relative gene expression levels due to the differences in factor composition. Our results indicate that whether an experimental design can provide all possible relative expression levels of all sample pairs for each gene should be the first criterion to be considered in an evaluation of experimental designs for two-color cDNA microarray experiments. 相似文献
3.
Ming Zhang Yudong Zhang Li Liu Lijuan Yu Shirley Tsang Jing Tan Wenhua Yao Manjit S Kang Yongqiang An Xingming Fan 《BMC bioinformatics》2010,11(1):1-16
Background
In the last decade, a large amount of microarray gene expression data has been accumulated in public repositories. Integrating and analyzing high-throughput gene expression data have become key activities for exploring gene functions, gene networks and biological pathways. Effectively utilizing these invaluable microarray data remains challenging due to a lack of powerful tools to integrate large-scale gene-expression information across diverse experiments and to search and visualize a large number of gene-expression data points.Results
Gene Expression Browser is a microarray data integration, management and processing system with web-based search and visualization functions. An innovative method has been developed to define a treatment over a control for every microarray experiment to standardize and make microarray data from different experiments homogeneous. In the browser, data are pre-processed offline and the resulting data points are visualized online with a 2-layer dynamic web display. Users can view all treatments over control that affect the expression of a selected gene via Gene View, and view all genes that change in a selected treatment over control via treatment over control View. Users can also check the changes of expression profiles of a set of either the treatments over control or genes via Slide View. In addition, the relationships between genes and treatments over control are computed according to gene expression ratio and are shown as co-responsive genes and co-regulation treatments over control.Conclusion
Gene Expression Browser is composed of a set of software tools, including a data extraction tool, a microarray data-management system, a data-annotation tool, a microarray data-processing pipeline, and a data search & visualization tool. The browser is deployed as a free public web service (http://www.ExpressionBrowser.com) that integrates 301 ATH1 gene microarray experiments from public data repositories (viz. the Gene Expression Omnibus repository at the National Center for Biotechnology Information and Nottingham Arabidopsis Stock Center). The set of Gene Expression Browser software tools can be easily applied to the large-scale expression data generated by other platforms and in other species. 相似文献4.
5.
Bushel PR Hamadeh H Bennett L Sieber S Martin K Nuwaysir EF Johnson K Reynolds K Paules RS Afshari CA 《Bioinformatics (Oxford, England)》2001,17(6):564-565
SUMMARY: MAPS is a MicroArray Project System for management and interpretation of microarray gene expression experiment information and data. Microarray project information is organized to track experiments and results that are: (1) validated by performing analysis on stored replicate gene expression data; and (2) queried according to the biological classifications of genes deposited on microarray chips. 相似文献
6.
Microarray analysis has become a key experimental tool in the study of genome‐wide patterns of gene expression. The labeling step of target molecules such as cDNA or cRNA plays a key role in a microarray experiment because the amount of mRNA is measured indirectly by the labeled molecules. In this paper, the most widely used cDNA labeling strategies in microarray experiments are reviewed in detail, including direct labeling and indirect labeling methods along with a discussion of the merits and disadvantages of these methods. Furthermore, various RNA amplification approaches were surveyed to obtain a target nucleic acid sufficient for microarray experiments from minute amounts of mRNA. Finally, the labeling strategies of commonly used microarray platforms (e.g., Affymetrix GeneChip®, CodeLink? Bioarray, Agilent and spotted microarrays) were compared. 相似文献
7.
Mandal D Srivastava A Mahlum E Desai D Maran A Yaszemski M Jalal SM Gitelis S Bertoni F Damron T Irwin R O'connor M Schwartz H Bolander ME Sarkar G 《Gene》2007,386(1-2):131-138
Deciphering the molecular basis of cancer is critical for developing novel diagnostic and therapeutic strategies. To better understand the early molecular events involving osteogenic sarcoma (OGS), we have initiated a program to identify potential tumor suppressor genes. Expression profiling of total RNA from ten normal bone cell lines and eleven OGS-derived cell lines by microarray showed 135-fold lower expression of FRZB/sFRP3 mRNA in OGS cells compared to bone cells; this down-regulation of Frzb/sFRP3 mRNA expression was found to be serum-independent. Subsequently, fourteen OGS biopsy specimens showed nine-fold down-regulation of Frzb/sFRP3 mRNA expression compared to expression in eight normal bone specimens as determined by microarray. FRZB /sFRP3 protein level was also found to be at a very low level in 4/4 OGS cell lines examined. Quantitation by RT-PCR indicated approximately 70% and approximately 90% loss of Frzb/sFRP3 mRNA expression in OGS biopsy specimens and OGS-derived cell lines respectively, compared to expression in bone (p<0.0001). Hybridization experiments of a cDNA microarray containing paired normal and tumor specimens from nineteen different organs did not show any significant difference in the level of Frzb/sFRP3 mRNA expression between the normal and the corresponding tumor tissues. Exogenous expression of FRZB/sFRP3 mRNA in two OGS-derived cell lines lacking endogenous expression of the mRNA produced abundant mRNA from the exogenous gene, eliminating degradation as a possibility for very low level of FRZB/sFRP3 mRNA in OGS specimens. Results from PCR-based experiments suggest that the FRZB/sFRP3 gene is not deleted in OGS cell lines, however, karyotyping shows gross abnormalities involving chromosome 2 (location of the FRZB gene) in five of twelve OGS-derived cell lines. Together, these data suggest a tumor-suppressive potential for FRZB/sFRP3 in OGS. 相似文献
8.
Two-color cDNA or oligonucleotide-based spotted microarrays have been commonly used in measuring the expression levels of thousands of genes simultaneously. To realize the immense potential of this powerful new technology, budgeted within limited resources or other constraints, practical designs with high efficiencies are in demand. In this study, we address the design issue concerning the arrangement of the mRNA samples labeled with fluorescent dyes and hybridized on the slides. A normalization model is proposed to characterize major sources of systematic variation in a two-color microarray experiment. This normalization model establishes a connection between designs for two-color microarray experiments with a particular class of classical row-column designs. A heuristic algorithm for constructing A-optimal or highly efficient designs is provided. Statistical optimality results are found for some of the designs generated from the algorithm. It is believed that the constructed designs are the best or very close to the best possible for estimating the relative gene expression levels among the mRNA samples of interest. 相似文献
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10.
Ricardo A. Verdugo Christian F. Deschepper Gloria Mu?oz Daniel Pomp Gary A. Churchill 《Nucleic acids research》2009,37(17):5610-5618
Measurements of gene expression from microarray experiments are highly dependent on experimental design. Systematic noise can be introduced into the data at numerous steps. On Illumina BeadChips, multiple samples are assayed in an ordered series of arrays. Two experiments were performed using the same samples but different hybridization designs. An experiment confounding genotype with BeadChip and treatment with array position was compared to another experiment in which these factors were randomized to BeadChip and array position. An ordinal effect of array position on intensity values was observed in both experiments. We demonstrate that there is increased rate of false-positive results in the confounded design and that attempts to correct for confounded effects by statistical modeling reduce power of detection for true differential expression. Simple analysis models without post hoc corrections provide the best results possible for a given experimental design. Normalization improved differential expression testing in both experiments but randomization was the most important factor for establishing accurate results. We conclude that lack of randomization cannot be corrected by normalization or by analytical methods. Proper randomization is essential for successful microarray experiments. 相似文献
11.
Background
MicroRNAs (miRNAs) are oligoribonucleotides with an important role in regulation of gene expression at the level of translation. Despite imperfect target complementarity, they can also significantly reduce mRNA levels. The validity of miRNA target gene predictions is difficult to assess at the protein level. We sought, therefore, to determine whether a general lowering of predicted target gene mRNA expression by endogenous miRNAs was detectable within microarray gene expression profiles. 相似文献12.
Performance comparison of one-color and two-color platforms within the MicroArray Quality Control (MAQC) project 总被引:18,自引:0,他引:18
Patterson TA Lobenhofer EK Fulmer-Smentek SB Collins PJ Chu TM Bao W Fang H Kawasaki ES Hager J Tikhonova IR Walker SJ Zhang L Hurban P de Longueville F Fuscoe JC Tong W Shi L Wolfinger RD 《Nature biotechnology》2006,24(9):1140-1150
Microarray-based expression profiling experiments typically use either a one-color or a two-color design to measure mRNA abundance. The validity of each approach has been amply demonstrated. Here we provide a simultaneous comparison of results from one- and two-color labeling designs, using two independent RNA samples from the Microarray Quality Control (MAQC) project, tested on each of three different microarray platforms. The data were evaluated in terms of reproducibility, specificity, sensitivity and accuracy to determine if the two approaches provide comparable results. For each of the three microarray platforms tested, the results show good agreement with high correlation coefficients and high concordance of differentially expressed gene lists within each platform. Cumulatively, these comparisons indicate that data quality is essentially equivalent between the one- and two-color approaches and strongly suggest that this variable need not be a primary factor in decisions regarding experimental microarray design. 相似文献
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14.
Fujibuchi W Kiseleva L Taniguchi T Harada H Horton P 《Bioinformatics (Oxford, England)》2007,23(22):3103-3104
The establishment and rapid expansion of microarray databases has created a need for new search tools. Here we present CellMontage, the first server for expression profile similarity search over a large database-69 000 microarray experiments derived from NCBI's; GEO site. CellMontage provides a novel, content-based search engine for accessing gene expression data. Microarray experiments with similar overall expression to a user-provided expression profile (e.g. microarray experiment) are computed and displayed-usually within 20 s. The core search engine software is downloadable from the site. 相似文献
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17.
The efficiency of pooling mRNA in microarray experiments 总被引:11,自引:0,他引:11
In a microarray experiment, messenger RNA samples are oftentimes pooled across subjects out of necessity, or in an effort to reduce the effect of biological variation. A basic problem in such experiments is to estimate the nominal expression levels of a large number of genes. Pooling samples will affect expression estimation, but the exact effects are not yet known as the approach has not been systematically studied in this context. We consider how mRNA pooling affects expression estimates by assessing the finite-sample performance of different estimators for designs with and without pooling. Conditions under which it is advantageous to pool mRNA are defined; and general properties of estimates from both pooled and non-pooled designs are derived under these conditions. A formula is given for the total number of subjects and arrays required in a pooled experiment to obtain gene expression estimates and confidence intervals comparable to those obtained from the no-pooling case. The formula demonstrates that by pooling a perhaps increased number of subjects, one can decrease the number of arrays required in an experiment without a loss of precision. The assumptions that facilitate derivation of this formula are considered using data from a quantitative real-time PCR experiment. The calculations are not specific to one particular method of quantifying gene expression as they assume only that a single, normalized, estimate of expression is obtained for each gene. As such, the results should be generally applicable to a number of technologies provided sufficient pre-processing and normalization methods are available and applied. 相似文献
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19.
Donatello Telesca Lurdes Y.T. Inoue Mauricio Neira Ruth Etzioni Martin Gleave Colleen Nelson 《Biometrics》2009,65(3):793-804
Summary . Time course microarray data consist of mRNA expression from a common set of genes collected at different time points. Such data are thought to reflect underlying biological processes developing over time. In this article, we propose a model that allows us to examine differential expression and gene network relationships using time course microarray data. We model each gene-expression profile as a random functional transformation of the scale, amplitude, and phase of a common curve. Inferences about the gene-specific amplitude parameters allow us to examine differential gene expression. Inferences about measures of functional similarity based on estimated time-transformation functions allow us to examine gene networks while accounting for features of the gene-expression profiles. We discuss applications to simulated data as well as to microarray data on prostate cancer progression. 相似文献
20.
Deutsch EW Ball CA Berman JJ Bova GS Brazma A Bumgarner RE Campbell D Causton HC Christiansen JH Daian F Dauga D Davidson DR Gimenez G Goo YA Grimmond S Henrich T Herrmann BG Johnson MH Korb M Mills JC Oudes AJ Parkinson HE Pascal LE Pollet N Quackenbush J Ramialison M Ringwald M Salgado D Sansone SA Sherlock G Stoeckert CJ Swedlow J Taylor RC Walashek L Warford A Wilkinson DG Zhou Y Zon LI Liu AY True LD 《Nature biotechnology》2008,26(3):305-312
One purpose of the biomedical literature is to report results in sufficient detail that the methods of data collection and analysis can be independently replicated and verified. Here we present reporting guidelines for gene expression localization experiments: the minimum information specification for in situ hybridization and immunohistochemistry experiments (MISFISHIE). MISFISHIE is modeled after the Minimum Information About a Microarray Experiment (MIAME) specification for microarray experiments. Both guidelines define what information should be reported without dictating a format for encoding that information. MISFISHIE describes six types of information to be provided for each experiment: experimental design, biomaterials and treatments, reporters, staining, imaging data and image characterizations. This specification has benefited the consortium within which it was developed and is expected to benefit the wider research community. We welcome feedback from the scientific community to help improve our proposal. 相似文献