RNA Biomarker Analysis

Quantitative Analysis of RNA Biomarkers

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

HER2 mRNA – ACD using DAB

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

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