Study to compare SET ER/PR index values from Affymetrix U133A microarrays to values from targeted RNASeq and to validate technical reproducibility on U133A microarrays
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ABSTRACT:
PROVIDER: PRJNA531737 | ENA |
REPOSITORIES: ENA
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Project description:63 samples were profiled in replicates on Affymetrix microarrays and using a custom RNASeq panel. The samples were randomly split in a training and validation cohort. SET ER/PR was calculated and a linear model was trained and validated to reproduce values obtained by microarray using RNASeq. In addition, the microarray replicates were used to validate the technical reproducibility of SET ER/PR.
2019-05-30 | GSE129560 | GEO
Project description:Recently, long oligonucleotide (60-70mer) microarrays for two-color experiments have been developed and are gaining widespread use. In addition, when there is limited availability of mRNA from tissue sources, RNA amplification can and is being used to produce sufficient quantities of cRNA for microarray hybridization. Taking advantage of the selective degradation of RNA under alkaline conditions, we have developed a method to “strip” glass-based oligonucleotide microarrays that use fluorescent RNA in the hybridization, while leaving the DNA oligonucleotide probes intact and usable for a second experiment. Replicate microarray experiments conducted using stripped arrays showed high reproducibility, however, we found that arrays could only be stripped and reused once without compromising data quality. The intraclass correlation (ICC) between a virgin array and a stripped array hybridized with the same sample showed a range of 0.90-0.98, which is comparable to the ICC of two virgin arrays hybridized with the same sample. Using this method, once stripped oligonucleotide microarrays are usable, reliable, and should help to reduce costs. Keywords = Agilent microarray Keywords = Stripped arrays Keywords = Replicate reproducibility Keywords: other
2005-02-23 | GSE1706 | GEO
Project description:In the teleost fish Fundulus heteroclitus there is extensive variation in gene expression among individuals within and between populations. Accurate measures of the variation in mRNA expression using microarrays can be confounded by technical variation, which includes variation in RNA isolation procedures, day of hybridization and methods used to amplify and dye label RNA for hybridization. In this manuscript we analyze the relationship between the amount of mRNA and the fluorescent signal from the microarray hybridizations demonstrating that for a wide-range of mRNA concentrations the fluorescent signal is a linear function of the amount of mRNA. Additionally, the separate isolation, labeling or hybridization of RNA does not add significant amounts of variation in microarray measures of gene expression. However, single or double rounds of amplification for labeling do have small but significant affects on 10% of genes, but this source of technical variation is easy to avoid. To examine both technical and stochastic biological variation, mRNA expression was measured from the same five individuals over a six-week time course. There were few, if any, meaningful differences in gene expression among time points. Thus, microarray measures using standard laboratory procedures can be precise and quantitative and are not subject to significant random biological noise. mRNA expression was measured using microarrays where each array had four spatially separated replicates per gene. The 384 F. heteroclitus cDNA microarrays were printed using 55 control genes and 329 cDNAs which encode essential proteins for cellular metabolism. To test for the relationship between fluorescence and the quantity of RNA, five concentrations of fluorescently labeled RNA were used: 1.2 to 700 pmol of Cy3 or Cy5. A 15 µl hybridization using the 384 cDNA array corresponds to 0.09 to 47 µM of Cy dye. Cy5 dye labeled RNA was used at concentrations 18% less than Cy3 because the Cy5 dye is a more efficient fluorophore (greater fluorescence per photon) than the Cy3 dye. The average of eight fluorescence values for each gene was normalized to the original concentration of RNA added.
2010-05-15 | E-GEOD-12858 | biostudies-arrayexpress
Project description:Four different concentrations (0, 0.75, 1.5, 3 pM) of genes were spiked in groups (group size 6-7 transcripts) Each concentration combination were spiked into a common backgroung sample (HeLa cell line) and hybridized to three HG-U133A Plus 2.0 Arrays. Keywords: Latin square design for HG-U133A Plus 2.0 GeneChips
2009-11-10 | GSE17968 | GEO
Project description:Technical replicates, consisting of four independent hybridizations of the same labelled cRNA, were performed to determine array-to-array reproducibility. After normalization, correlation index between all replicates exceeded 98%. In this study, we analyzed reproducibility of Agilent-016251 Sparus aurata Oligo Microarray based on single-colour detection (Cyanine-3 only). A RNA sample, prepared mixing four different larval stages of gilthead sea bream, was labelled in according to Agilent One-Color Microarray-Based Gene Expression Analysis protocol. The same labelled cRNA was used for four independent fragmentation reactions and then hybridized spanning all array positions of 4x44K platform. Microarrays are scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides are scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software creates a unique ID for each pair of XDR scans and saves it to both scan image files. Feature Extraction 9.5 uses XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended Background-Subtracted Signal.
2009-02-19 | E-GEOD-12118 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE12858: Technical Analysis of Fundulus heteroclitus cDNA Microarrays, experiment A GSE12898: Technical Analysis of Fundulus heteroclitus cDNA Microarrays, experiment B Refer to individual Series
2010-05-15 | E-GEOD-13687 | biostudies-arrayexpress
Project description:Technical replicates, consisting of four independent hybridizations of the same labelled cRNA, were performed to determine array-to-array reproducibility. After normalization, correlation index between all replicates exceeded 98%.
2009-02-20 | GSE12118 | GEO
Project description:In the teleost fish Fundulus heteroclitus there is extensive variation in gene expression among individuals within and between populations. Accurate measures of the variation in mRNA expression using microarrays can be confounded by technical variation, which includes variation in RNA isolation procedures, day of hybridization and methods used to amplify and dye label RNA for hybridization. In this manuscript we analyze the relationship between the amount of mRNA and the fluorescent signal from the microarray hybridizations demonstrating that for a wide-range of mRNA concentrations the fluorescent signal is a linear function of the amount of mRNA. Additionally, the separate isolation, labeling or hybridization of RNA does not add significant amounts of variation in microarray measures of gene expression. However, single or double rounds of amplification for labeling do have small but significant affects on 10% of genes, but this source of technical variation is easy to avoid. To examine both technical and stochastic biological variation, mRNA expression was measured from the same five individuals over a six-week time course. There were few, if any, meaningful differences in gene expression among time points. Thus, microarray measures using standard laboratory procedures can be precise and quantitative and are not subject to significant random biological noise.
2008-11-21 | GSE12858 | GEO
Project description:Both spotted long oligonucleotide arrays (GPL1384) and Affymetrix GeneChip arrays (GPL96) were used to analyze gene expression in six human head and neck squamous cell carinoma samples versus control samples or lymph node metastases of the same patients. Hybridizations of HG-U133A GeneChip arrays were performed using standard Affymetrix protocols and equipment. Before hybridization on DKFZ Operon 27k long oligonucleotide arrays, 2 µg RNA were amplified by one round of linear isothermal RNA amplification, followed by Cy-dUTP incorporation using Klenow fragment. Hybridizations were performed for 16 h at 42 °C in a GeneTAC Hybridization Station (Genomic Solutions) using UltraHyb hybridization buffer (Ambion). Hybridized microarrays were scanned at 5 µm resolution on a GenePix 4000B microarray scanner (Axon Instruments). Raw signal intensities from both platforms were normalized applying variance stabilization (W. Huber et al., Bioinformatics 18 Suppl 1, 2002). Expression ratios were compared for those genes represented in both array platforms. Keywords: parallel sample
2005-03-01 | GSE1722 | GEO
Project description:Worms spun at higher than normal g values for 4 days on 20G centrifuge vs. worms kept at 1 X G. Both are mixed stage cultures. A stimulus or stress experiment design type is where that tests response of an organism(s) to stress/stimulus. e.g. osmotic stress, behavioral treatment Physical Characteristics: worms kept at increased g values for 4 days (x g value)
2013-06-22 | GSE38877 | GEO