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

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 PAX-gene blood RNA tube is sufficient for downstream microarray analysis. Copyright © American Society for Investigative Pathology and the Association for Molecular Pathology.