title: 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 PAXgene blood RNA tube is sufficient for downstream microarray analysis.
authors: Brendstrupgaardsvej 102, DK-8200 Aarhus N, Denmark. mkruhoffer@arosab.com
authors: Mogens
authors: M
authors: Kruhøffer
authors: Lars
authors: L
authors: Dyrskjøt
authors: Thorsten
authors: T
authors: Voss
authors: Raija L P
authors: RL
authors: Lindberg
authors: Ralf
authors: R
authors: Wyrich
authors: Thomas
authors: T
authors: Thykjaer
authors: Torben F
authors: TF
authors: Orntoft
mesh_terms: 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
chemical_list: D035683:MicroRNAs; D004247:DNA
references: Oncogene. 2006 Apr 20;25(17):2537-45
references: 16331254
references: Oncogene. 2006 Oct 9;25(46):6170-5
references: 17028596
references: Curr Mol Med. 2006 May;6(3):339-49
references: 16712479
references: Rheumatology (Oxford). 2006 Jun;45(6):694-702
references: 16418202
references: Expert Rev Mol Diagn. 2006 Nov;6(6):843-55
references: 17140371
references: Proc Natl Acad Sci U S A. 2007 Feb 20;104(8):2750-5
references: 17293455
references: Pharmacogenet Genomics. 2005 Jul;15(7):483-92
references: 15970796
references: J Immunol Methods. 2002 Mar 1;261(1-2):195-7
references: 11861077
references: Clin Chem. 2002 Nov;48(11):1883-90
references: 12406972
references: Leukemia. 2002 Dec;16(12):2395-9
references: 12454744
references: Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1896-901
references: 12578971
references: Pharmacogenomics J. 2004;4(3):193-207
references: 15037859
references: Cancer Res. 2004 Jun 1;64(11):3753-6
references: 15172979
references: Cancer Chemother Pharmacol. 2004 Sep;54(3):265-72
references: 15133628
references: Clin Biochem. 2004 Sep;37(9):741-4
references: 15329310
references: Physiol Genomics. 2004 Nov 17;19(3):247-54
references: 15548831
references: J Nutr. 2005 Jan;135(1):42-7
references: 15623830
references: Drug Discov Today. 2005 Jul 15;10(14):965-76
references: 16023055
references: Cancer Res. 2005 Aug 15;65(16):7065-70
references: 16103053
references: Breast Cancer Res. 2005;7(5):R634-44
references: 16168108
references: Cell Cycle. 2005 Sep;4(9):1179-84
references: 16096373
references: Development. 2005 Nov;132(21):4653-62
references: 16224045
references: Atherosclerosis. 2006 Mar;185(1):97-107
references: 16002074
references: Future Oncol. 2006 Feb;2(1):73-82
references: 16556074
references: BMC Mol Biol. 2006;7:3
references: 16448564
references: Physiol Genomics. 2006 Apr 13;25(2):203-15
references: 16403844
references: Mol Med. 2005 Jan-Dec;11(1-12):21-9
references: 16622521
references: Nat Rev Cancer. 2006 Jul;6(7):565-71
references: 16794639
references: Cancer Res. 2006 Aug 1;66(15):7390-4
references: 16885332
references: RNA. 2006 May;12(5):913-20
references: 16540696