Quantitative Analysis of RNA Biomarkers

Quantitative Analysis of RNA Biomarkers

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Quantitative Measurement of mRNA in Addition to QIF of Proteins to Improve Risk Assessment and Predictive Value of Companion Diagnostics

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HER2 mRNA – ACD using DAB

Img: Her2 mRNA

HER2 mRNA Spot 190 (block B): Score=4012

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miRNA In situ Hybridization

  • Provide direct assessment of abundance and spatial expression of miRNAs
  • LNA modified oligonucleotide probes
    • Low Tm of duplex since only ~20mer probes
    • LNA = “Locked Nucleic Acid”
    • Methylene bridge between 2’-O and 4’-C locks sugar in N-type conformation
    • Increased thermodynamic stability of duplex providing increased Tm for greater specificity
  • DIG Conjugated Probes
  • Proteinase K, 4% formaldehyde & EDC to fix RNAs
  • Multiplexed with Cytokeratin and Dapi for Compartmentalization and AQUA Analysis

Img: hybridization

Spot 190

Img: Spot 190

Breast Cancer

Img: breast cancer