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J Mol Diagn. 2007 Sep;9(4):452-8. Epub 2007 Aug 9.

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

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1
Brendstrupgaardsvej 102, DK-8200 Aarhus N, Denmark. mkruhoffer@arosab.com

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.

PMID:
17690207
PMCID:
PMC1975097
DOI:
10.2353/jmoldx.2007.060175
[Indexed for MEDLINE]
Free PMC Article
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