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Belbin TJ Gaspar J Haigentz M Perez-Soler R Keller SM Prystowsky MB Childs G Socci ND 《BioTechniques》2004,36(2):310-314
The use of universal RNA reference sets is an increasingly common approach to molecular classification studies with cDNA microarrays. Here we evaluated the reliability of indirect measurements of fluorescence ratios with a common RNA reference as a means of identifying differentially expressed genes. Comparisons of direct and indirect measures of differential gene expression showed a strong overall correlation in fluorescence ratio measurements but also a high degree of false positives in our indirect measurements. These results indicated that the application of more stringent ratio filters may be required when assessing differential gene expression utilizing a common RNA reference in classification studies. 相似文献
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We used scaled factorial moments to search for intermittency in the log expression ratios (LERs) for thousands of genes spotted on cDNA microarrays (gene chips). Results indicate varying levels of intermittency in gene expression. The observation of intermittency in the data analyzed provides a complimentary handle on moderately expressed genes, generally not tackled by conventional techniques. 相似文献
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Monitoring gene expression using DNA microarrays 总被引:13,自引:0,他引:13
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Background
One of the primary tasks in analysing gene expression data is finding genes that are differentially expressed in different samples. Multiple testing issues due to the thousands of tests run make some of the more popular methods for doing this problematic. 相似文献9.
Global analysis of gene expression using GeneChip microarrays 总被引:13,自引:0,他引:13
Zhu T 《Current opinion in plant biology》2003,6(5):418-425
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As the data generated by microarray technology continue to amass, it is necessary to compare and combine gene expression data from different platforms. To evaluate the performance of cDNA and long oligonucleotide (60-mer) arrays, we generated gene expression profiles for two cancer cell lines and compared the data between the two platforms. All 6182 unique genes represented on both platforms were included in the analysis. A limited correlation (r = 0.4708) was obtained and the difference in measurement of low-expression genes was considered to contribute to the limited correlation. Further restriction of the data set to differentially expressed genes detected in cDNA microarrays (1205 genes) and oligonucleotide arrays (1325 genes) showed modest correlations of 0.7076 and 0.6441 between the two platforms. Quantitative real-time PCR measurements of a set of 10 genes showed better correlation with oligonucleotide arrays. Our results demonstrate that there is substantial variation in the data generated from cDNA and 60-mer oligonucleotide arrays. Although general agreement was observed in measurements of differentially expressed genes, we suggest that data from different platforms could not be directly amassed. 相似文献
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This study established the utility of cross-species application of the cDNA microarray technique for investigating differential gene expression. Using both total RNA and mRNA samples recovered from two opossum cell lines derived from UVB-induced melanoma, we analyzed expression of ca. 4400 genes on the human DermArray DNA microarrays. The signals generated on the DermArrays were clear, strong, and reproducible. A cDNA dot blot consisting of differentially expressed genes representative of different functional clusters was used to validate the DermArray results. We also cloned a Monodelphis gene, keratin 18 (KRT18), and characterized its expression patterns in tumor samples of different progression stages. Up-regulated expression was observed for the KRT18 gene in advanced melanomas, a finding consistent with the DermArray analysis. These results provide evidence that cross-species application of cDNA microarrays is a useful strategy for investigating gene expression patterns in animal models for which species-specific cDNA microarrays are not available. 相似文献
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Towards
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genome analysis: monitoring expression profiles of 1400 genes using cDNA microarrays 总被引:16,自引:3,他引:13
Yijun Ruan James Gilmore Timothy Conner 《The Plant journal : for cell and molecular biology》1998,15(6):821-833
cDNA microarrays containing 1443 Arabidopsis thaliana genes were analyzed for expression profiles in major organs of Arabidopsis plants. Novel expression profiles were identified for many coding sequences with putative gene identifications. Expression patterns of novel sequences provided clues to their possible functions. The results demonstrate how microarrays containing a large number of Arabidopsis genes can provide a powerful tool for plant gene discovery, functional analysis and elucidation of genetic regulatory networks. 相似文献
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We propose a statistical model for estimating gene expression using data from multiple laser scans at different settings of hybridized microarrays. A functional regression model is used, based on a non-linear relationship with both additive and multiplicative error terms. The function is derived as the expected value of a pixel, given that values are censored at 65 535, the maximum detectable intensity for double precision scanning software. Maximum likelihood estimation based on a Cauchy distribution is used to fit the model, which is able to estimate gene expressions taking account of outliers and the systematic bias caused by signal censoring of highly expressed genes. We have applied the method to experimental data. Simulation studies suggest that the model can estimate the true gene expression with negligible bias. AVAILABILITY: FORTRAN 90 code for implementing the method can be obtained from the authors. 相似文献
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Quantitative assessment of filter-based cDNA microarrays: gene expression profiles of human T-lymphoma cell lines 总被引:1,自引:0,他引:1
MOTIVATION: While the use of cDNA microarrays for functional genomic analysis has become commonplace, relatively little attention has been placed on false positives, i.e. the likelihood that a change in measured radioactive or fluorescence intensity may reflect a change in gene expression when, in fact, there is none. Since cDNA arrays are being increasingly used to rapidly distinguish biomarkers for disease detection and subsequent assay development (Wellman et al., Blood, 96, 398-404, 2000), the impact of false positives can be significant. For the use of this technology, it is necessary to develop quantitative criteria for reduction of false positives with radioactively-labeled cDNA arrays. RESULTS: We used a single source of RNA (HuT78 T lymphoma cells) to eliminate sample variation and quantitatively examined intensity ratios using radioactively labeled cDNA microarrays. Variation in intensity ratios was reduced by processing microarrays in side-by-side (parallel mode) rather than by using the same microarray for two hybridizations (sequential mode). Based on statistical independence, calculation of the expected number of false positives as a function of threshold showed that a detection limit of [log(2)R] >0.65 with agreement from three replicates could be used to identify up- or down-modulated genes. Using this quantitative criteria, gene expression differences between two related T lymphoma cell lines, HuT78 and H9, were identified. The relevance of these findings to the known functional differences between these cell types is discussed. 相似文献
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Reliability of gene expression ratios for cDNA microarrays in multiconditional experiments with a reference design 总被引:3,自引:0,他引:3
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In a typical gene expression profiling experiment with multiple conditions, a common reference sample is used for co-hybridization with the samples to yield expression ratios. Differential expression for any other sample pair can then be calculated by assembling the ratios from their hybridizations with the reference. In this study we test the validity of this approach. Differential expression of a sample pair (i, j) was obtained in two ways: directly, by hybridizations of sample i versus j, and indirectly, by multiplying the expression ratios for hybridizations of sample i versus pool and pool versus sample j. We performed gene expression profiling using amphibian embryos (Xenopus laevis). Every sample combination of four different stages and a pool was profiled. Direct and indirect values were compared and used as the quality criterion for the data. Based on this criterion, 82% of all ratios were found to be sufficiently accurate. To increase the reliability of the signals, several widely used filtering techniques were tested. Filtering by differences of repeated hybridizations was found to be the optimal filter. Finally, we compared microarray-based gene expression profiles with the corresponding expression patterns obtained by whole-mount in situ hybridizations, resulting in a 90% correspondence. 相似文献
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