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Originally published online as doi:10.2353/jmoldx.2007.060175 on August 9, 2007

Published online before print August 9, 2007
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Journal of Molecular Diagnostics 2007, Vol. 9, No. 4
Copyright © 2007 American Society for Investigative Pathology & Association for Molecular Pathology
DOI: 10.2353/jmoldx.2007.060175

Technical Advance

Isolation of Microarray-Grade Total RNA, MicroRNA, and DNA from a Single PAXgene Blood RNA Tube

Mogens Kruhøffer*{dagger}, Lars Dyrskjøt*, Thorsten Voss{ddagger}, Raija L.P. Lindberg§, Ralf Wyrich{ddagger}, Thomas Thykjaer{dagger} and Torben F. Orntoft*{dagger}

From the Molecular Diagnostic Laboratory, * Clinical Biochemical Department, Aarhus University Hospital, Aarhus, Denmark; AROS Applied Biotechnology A/S, {dagger} Aarhus, Denmark; Qiagen, {ddagger} Hilden, Germany; and the Clinical Neuroimmunology Laboratory, § Departments of Research and Neurology, University Hospital Basel, Basel, Switzerland

Abstract

We have developed a procedure for isolation of microRNA and genomic DNA in addition to total RNA from whole blood stabilized in PAXgene Blood RNA tubes. The procedure is based on automatic extraction on a BioRobot MDx and includes isolation of DNA from a fraction of the stabilized blood and recovery of small RNA species that are otherwise lost. The procedure presented here is suitable for large-scale experiments and is amenable to further automation. Procured total RNA and DNA was tested using Affymetrix Expression and single-nucleotide polymorphism GeneChips, respectively, and isolated microRNA was tested using spotted locked nucleic acid-based microarrays. We conclude that the yield and quality of total RNA, microRNA, and DNA from a single PAXgene blood RNA tube is sufficient for downstream microarray analysis.






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Copyright © 2007 by the American Society for Investigative Pathology and the Association for Molecular Pathology.