HTG Molecular’s qNPA technology allows laboratories to quickly and accurately measure gene expression levels in a wide range of sample types, including formalin fixed paraffin embedded (FFPE) tissue. Meaningful data can be obtained from very small, highly degraded, or archived FFPE tissue. The simplified qNPA protocol requires no RNA extraction, no RNA amplification, and no RNA labeling, removing the manipulations that introduce variability inherent in in other technologies.

• Laboratories can use virtually any specimen type, whether it is cultured cells, fresh/frozen tissue, or FFPE. For FFPE samples, simply scrape the entire section off the slide into a vial, or choose to microdissect just the tumor area of the section to enhance the specificity of the results.
• The qNPA reaction takes place in the sample prep vial. HTG Molecular’s proprietary lysis buffer with gene-specific probes is added to the vial. The lysis buffer opens the cells and allows the probes to bind to the target RNA.
• All nucleic acids that are not bound to the probes are removed with the S1 Nuclease step — effectively enriching for targeted RNA — while removing all non-targeted RNAs, and excess qNPA probes.
• At this point the reaction is halted with a stop solution, which destroys the S1 nuclease and hydrolyzes the bound target RNA. The solution is then neutralized and left with probes representative of the native RNA.
• The rest of the process is simple detection and imaging on the array plate and HTG Molecular’s ^SuperCAPELLA imaging system. Data from the imager is then delivered to the customer for normalization and analysis.