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TaqMan Gene Expression Arrays for Accurate Quantification of Toxicity Targets

TaqMan Gene Expression Arrays for Accurate Quantification of Toxicity Targets. Agenda. Test Procedure Specifications Validation Results. Test Procedure. RNA isolation cDNA generation (may be skipped with one-step RT-PCR) Card loading and sealing Thermal cycling Data Analysis.

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TaqMan Gene Expression Arrays for Accurate Quantification of Toxicity Targets

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  1. TaqMan Gene Expression Arrays for Accurate Quantification of Toxicity Targets

  2. Agenda • Test Procedure • Specifications • Validation Results

  3. Test Procedure • RNA isolation • cDNA generation (may be skipped with one-step RT-PCR) • Card loading and sealing • Thermal cycling • Data Analysis

  4. Dynamic Range in the Cytokine Card

  5. Dynamic Range in the Cytokine Card

  6. 3 cytokine cards with 20 plasmid targets Calculate dCt Calibrator Calculate dCT sample 1 Calculate dCt sample 2 Calculate RelQuant Values (20 for each sample--40 total) Normalize RQ values to positive fold Calculate Average of 40 RQ values Average plus 6 Stdev is Fold Discrimination Fold-Discrimination in the Cytokine Card

  7. Specifications: Relative Quantification (median/ttest) for 3 Cytokine Cards Avg 1.34 Stdev 0.41 5.00 4.50 Spec 4.00 Avg + 6xStdev = 3.79 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00 0 5 10 15 20 25 30 35 40

  8. DNAx: Cytokine Gene Expression and PBMC Activation

  9. Source Precision Medicine: Cytokine Gene Expression and PBMC Activation

  10. Phase 1 Molecular Toxicology: Cytokine Gene Expression in Liver Cell Cultures

  11. Phase 1 Molecular Toxicology: MMC Incubations of Liver Cells in the Human Toxicology Card

  12. Phase 1 Molecular Toxicology: MMS Incubations of Liver Cells in the Human Toxicology Card

  13. Phase 1 Molecular Toxicology: Carnustine Incubations of Liver Cells in the Human Toxicology Card

  14. Phase 1 Molecular Toxicology: Cytokine Gene Expression in RNA Isolated from Blood Cells

  15. Phase 1 Molecular Toxicology: Toxicology Gene Expression in RNA Isolated from Blood Cells

  16. Phase 1 Molecular Toxicology: Cytokine Gene Expression in RNA Isolated from Lymphocyte Cultures

  17. Phase 1 Molecular Toxicology: Toxicology Gene Expression in RNA Isolated from Lymphocyte Cultures

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