Mogens Kruhøffer, Lars Dyrskjøt, Thorsten Voss, Raija L. P

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Presentation transcript:

Isolation of Microarray-Grade Total RNA, MicroRNA, and DNA from a Single PAXgene Blood RNA Tube Mogens Kruhøffer, Lars Dyrskjøt, Thorsten Voss, Raija L.P. Lindberg, Ralf Wyrich, Thomas Thykjaer, Torben F. Orntoft The Journal of Molecular Diagnostics Volume 9, Issue 4, Pages 452-458 (September 2007) DOI: 10.2353/jmoldx.2007.060175 Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Overview scheme for the TRI-X workflow. TRI-X is a combination of three methods, starting with 2.5-ml whole blood collected in a single PAXgene Blood RNA tube. Genomic DNA is isolated from a 1-ml aliquot with the QIAamp Blood DNA protocol. The remaining sample is extracted with the PAXgene Blood RNA MDx procedure to isolate mRNAs and large rRNA fragments. Small RNA species including miRNA are isolated from the flow through of the PAXgene Blood RNA MDx protocol binding step via a modified RNeasy procedure. The Journal of Molecular Diagnostics 2007 9, 452-458DOI: (10.2353/jmoldx.2007.060175) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Sample mismatch analysis showing IBS scores of chip to chip comparisons of 250K SNP array. Heat map showing the result of a sample mismatch analysis comparing all SNP arrays against each other. The color of each square indicates the IBS score between SNP samples ranging from 0.9 to 2. The gray boxes indicate the average IBS score and SD between each group. Cnt, control DNA. The Journal of Molecular Diagnostics 2007 9, 452-458DOI: (10.2353/jmoldx.2007.060175) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Electropherograms from Agilent Bioanalyzer. The y axis represents fluorescence units [FU], and the x axis represents the runtime (s). A: Typical picture of a total RNA preparation with the PAXgene Blood RNA MDx protocol. The bands of the 18S and 28S rRNA fragments are clearly visible, and the RNA integrity number score is 8.7. Only very limited amounts of small RNA species are visible. B: Typical picture of a small RNA preparation isolated from the flow through from the PAXgene Blood RNA MDx binding step. Small RNAs including miRNA are clearly enriched. The Journal of Molecular Diagnostics 2007 9, 452-458DOI: (10.2353/jmoldx.2007.060175) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions