Literature DB >> 17690207

Isolation of microarray-grade total RNA, microRNA, and DNA from a single PAXgene blood RNA tube.

Mogens Kruhøffer1, Lars Dyrskjøt, Thorsten Voss, Raija L P Lindberg, Ralf Wyrich, Thomas Thykjaer, Torben F Orntoft.   

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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Substances：

Year:  2007        PMID： 17690207      PMCID： PMC1975097          DOI： 10.2353/jmoldx.2007.060175

Source DB:  PubMed          Journal:  J Mol Diagn        ISSN： 1525-1578            Impact factor:   5.568


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