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RNA Stabilization of Peripheral Blood and Profiling by Bead Chip Analysis

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DNA and RNA Profiling in Human Blood

Part of the book series: METHODS IN MOLECULAR BIOLOGY™ ((MIMB,volume 496))

Abstract

Gene expression profiling of peripheral blood is a very attractive approach for the development of new diagnostic markers of blood-borne but also systemic diseases as well as the development of biomarkers for drug development. Since most cellular components of peripheral blood are specialized to quickly respond to exogenous stimuli, sample procurement approaches are required that reduce the overall impact of ex vivo changes in gene expression due to technical issues such as prolonged sample handling or temperature changes during transportation of the blood prior to genome-wide analysis. To address these needs, a whole blood RNA stabilization technology was combined with a bead-based oligonucleotide microarray technology for genome-wide transcriptome analysis. Cells, and thereby also RNA is immediately stabilized after the blood draw by a commercially available device (PAXgene). Total RNA is then extracted from PAXgene-stabilized blood and subjected to microarray analysis. In our hands, the Illumina BeadChip array platform outperformed other microarray platforms. Combining RNA stabilization of peripheral blood with bead-based oligonucleotide microarray technology is not only applicable to small single-center studies with optimized infrastructure but also to large scale multi-center trials that are mandatory for the development of predictive markers for disease and treatment outcome.

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Acknowledgment

This work was mainly supported by the Alexander von Humboldt Foundation via a Sofja-Kovalevskaja Award to JLS, JLS is a member of the National Genome Research Network (NGFN) in Germany.

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Authors and Affiliations

  1. Department for Genomics, Life and Medical Sciences, University of Bonn, Bonn, Germany

    Svenja Debey-Pascher, Daniela Eggle & Joachim L. Schultze

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  1. Svenja Debey-Pascher
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  2. Daniela Eggle
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  3. Joachim L. Schultze
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Editors and Affiliations

  1. Institute of Transfusion Medicine and Immunology, Heidelberg University Medical Faculty of Mannheim, Mannheim, Germany

    Peter Bugert

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Debey-Pascher, S., Eggle, D., Schultze, J.L. (2009). RNA Stabilization of Peripheral Blood and Profiling by Bead Chip Analysis. In: Bugert, P. (eds) DNA and RNA Profiling in Human Blood. METHODS IN MOLECULAR BIOLOGY™, vol 496. Humana Press. https://doi.org/10.1007/978-1-59745-553-4_13

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  • DOI: https://doi.org/10.1007/978-1-59745-553-4_13

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-934115-93-0

  • Online ISBN: 978-1-59745-553-4

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