Dataset Information

Microarray probes and probe sets.

ABSTRACT: DNA microarrays have gained wide use in biomedical research by simultaneously monitoring the expression levels of a large number of genes. The successful implementation of DNA microarray technologies requires the development of methods and techniques for the fabrication of microarrays, the selection of probes to represent genes, the quantification of hybridization, and data analysis. In this paper, we concentrate on probes that are either spotted or synthesized on the glass slides through several aspects: sources of probes, the criteria for selecting probes, tools available for probe selections, and probes used in commercial microarray chips. We then provide a detailed review of one type of DNA microarray: Affymetrix GeneChips, discuss the need to re-annotate probes, review different methods for regrouping probes into probe sets, and compare various redefinitions through public available datasets.

SUBMITTER: Liu H

PROVIDER: S-EPMC3902802 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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Microarray probes and probe sets.

Liu Hongfang H Bebu Ionut I Li Xin X

Frontiers in bioscience (Elite edition) 20100101 1

DNA microarrays have gained wide use in biomedical research by simultaneously monitoring the expression levels of a large number of genes. The successful implementation of DNA microarray technologies requires the development of methods and techniques for the fabrication of microarrays, the selection of probes to represent genes, the quantification of hybridization, and data analysis. In this paper, we concentrate on probes that are either spotted or synthesized on the glass slides through severa ...[more]

PMID: 20036881

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Project description:MicroRNAs (miRNAs) have been shown to play an important role in many different cellular, developmental, and physiological processes. Accordingly, numerous methods have been established to identify and quantify miRNAs. The shortness of miRNA sequence results in a high dynamic range of melting temperatures and, moreover, impedes a proper selection of detection probes or optimized PCR primers. While miRNA microarrays allow for massive parallel and accurate relative measurement of all known miRNAs, they have so far been less useful as an assay for absolute quantification. Here, we present a microarray based approach for global and absolute quantification of miRNAs. The method relies on an equimolar pool of about 1000 synthetic miRNAs of known concentration which is used as an universal reference and labeled and hybridized in a dual colour approach on the same array as the sample of interest. Each single miRNA is quantified with respect to the universal reference outbalancing bias related to sequence, labeling, hybridization or signal detection method. We demonstrate the accuracy of the method by various spike in experiments. Further, we quantified miRNA copy numbers in liver samples and CD34(+)CD133(-) hematopoietic stem cells. Different probe designs were investigated with respect to sensitivity and selectivity of microarray hybridization. An array with 11 different variants of oligonucleotides for miR-122a and miR-16 was produced. 5 µg of respective total RNA was fluorescently labelled by 3’ ligation. Total RNA was hybridized in a dual colour approach to microarrays versus a second labelled synthetic miRNA pool. The synthetic miRNA pool consisted of 5 fmol of each of 816 non redundant miRNAs sequences and miRControl 3 sequences. Local background was subtracted from the signal to obtain the net signal intensity and the mean of the net signal intensities of 4 corresponding spots representing the same miRNA was computed for those spots only which were unflagged (empty spots, poor spots, negative spots) and for which the fluorescent intensity of the miRNAs derived from the samples of interest was two-fold the mean background value (2bkg dataset).

Dataset Information

Microarray probes and probe sets.

Publications

Microarray probes and probe sets.

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