Isolation of microarray-grade total RNA, microRNA, and DNA from a single PAXgene blood RNA tube.
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.
Kruhøffer
Mogens
M
Brendstrupgaardsvej 102, DK-8200 Aarhus N, Denmark. mkruhoffer@arosab.com
Dyrskjøt
Lars
L
Voss
Thorsten
T
Lindberg
Raija L P
RL
Wyrich
Ralf
R
Thykjaer
Thomas
T
Orntoft
Torben F
TF
D001800:Blood Specimen Collection / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D013997:Time Factors
D035683:MicroRNAs; D004247:DNA
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