Probe Hybridization and Signal Amplification in RNA In Situ Hybridization: A Technique for Detecting Specific RNA Sequences in Tissue Sections

To start hybridization, tap and flick the slides to remove any excess liquid and place them back in the slide rack. Remove prewarmed HPV probe from the oven and add approximately four drops to entirely cover each section. Cover the tray with a lid and insert it into the oven for 2 hours at 40 degrees Celsius. After completing incubation, remove the tray from the oven and remove the slide rack.

One slide at a time, quickly remove any excess liquid and place the slide in a slide rack submerged in a staining dish filled with 1x wash buffer, washing the slides for 2 minutes at room temperature with constant agitation, and repeat with fresh 1x wash buffer. Tap and flick to remove any excess liquid from the slides and place them in the slide rack again. Add approximately four drops of room temperature AMP1 per section to entirely cover each section. Cover the tray with a lid and insert it into the oven for 30 minutes at 40 degrees Celsius.

After removing the tray from the oven, remove the slide rack. Working one slide at a time, quickly remove any excess liquid and place the slide in the slide rack submerged in a staining dish filled with 1x wash buffer, washing for 2 minutes at room temperature with constant agitation, and repeat with fresh 1x wash buffer.