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RNAscope® VS Duplex Assay

The RNAscope® VS Duplex Assay is designed as an automated assay on the Discovery Ultra Automated IHC/ISH slide staining systems from Roche. The RNAscope® VS Duplex Assay, is an RNA in situ hybridization assay based on ACD’s patented signal amplification and background suppression technology. The assay uses a novel and proprietary method of in situ hybridization (ISH) to visualize single RNA molecules per cell in a multitude of sample types mounted on slides. RNAscope® Assays do not require the RNA-free environment used for traditional ISH.

RNAscope® VS Duplex Kit is designed for simultaneous in situ detection of two RNA species. 

Common applications include co-localization studies to map co-expression of two targets within the same cellular context (e.g. secreted ligand and its receptor) or to profile gene expression in a specific cell type expressing a known marker (e.g. a specific stem cell marker). To distinguish between the two targets, individual probes are used in probe channels 1 (C1) and Channel 2 ( C2). Channel 1 (C1)  is detected using HRP based diaminobenzidine (DAB) reaction resulting in a dark brown signal and Channel 2 (C2)  is detected using an AP Fast Red based reaction resulting in a red color. Due to the brightness of the red chromogen, it is advised that the lower expressed target be ordered in the C2 channel and stronger expressed target be ordered in the C1 channel.The stained slides are visualized with a bright-field microscope. 

The RNAscope® VS Duplex Reagent Kit automated workflow requires the following from Ventana Medical System  (

CAT #Order CodeProduct Name and Description
760-601408127115001mRNA Link
760-24908127174001mRNA Duplex Amp Kit
760-24808127166001mRNA Sample Prep Kit
960-761 — 960-785 250 Test Probe Dispenser User-Fillables 1-25
760-23407099037001mRNA RED Detection Kit
760-22406614353001mRNA DAB Detection Kit


The RNAscope® VS Duplex Reagent Kit automated workflow requires the following from ACD:

  • RNAscope®  2.5 VS Target Probes (Catalog or Made-to-Order VS Probes)
  • RNAscope® 2.5 VS Duplex Control Probes (Select species-specific VS positive control probes or packs of probes and VS negative control probes)
  • RNAscope® Control Slides (optional)
  • RNAscope® VS Duplex Reagent Kit
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Advanced Cell Diagnostics

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