Detection
| HTG Molecular’s detection steps described here capture and measure probe quantities representative of the native RNA as generated by the qNPA process.
The universal 96-well array plate is manufactured with generic anchor linker sequences to enable sensitive and specific measurement of each qNPA probe. |
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- The universal array plate is hybridized with programming linker probes, creating gene-specific hybridization spots which will subsequently bind and hybridize with the probes from the qNPA lysate.
- Gene-specific detection linker probes are added and hybridize to the qNPA probes.
- Universal detection probes are added, which bind to all available probe/linker combinations that are bound to anchors on the plate. Higher quantities of bound detection probes result in a stronger gene expression signal.
- A Streptavidin -horseradish peroxidase (SA-HRP) conjugate is added and binds to the detection probes, A chemiluminescent substrate is then introduced to the array plate. This combination produces light photons which are counted and interpreted as relative gene expression levels or relative light units on the SuperCAPELLA imaging system.
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