SEERNA® FISH RNA Fluorescence In Situ Hybridization Kit

The SEERNA® FISH RNA Fluorescence In Situ Hybridization Kit enables customized probe design targeting mRNA, lncRNA, and other RNA molecules for validation of RNA expression levels and spatial localization.
This kit allows simultaneous validation of 1–4 targets in a single experiment, providing a research tool with subcellular resolution, high sensitivity, and high specificity.
It is particularly suitable for validating the spatial expression of key targets or marker molecules identified through NGS or scRNA-seq analyses. It is especially useful for RNA targets lacking effective antibodies and for secreted proteins. The kit can also be applied to tissue detection of exogenous infectious viruses such as HIV, HCV, and HPV. In drug development, it supports preclinical drug target validation and drug toxicity evaluation.

Kit Components
· Buffer Reagent Kit (A): For sample pretreatment and reaction buffers; storage at 2–8°C
· Reaction Reagent Kit (B): Contains reaction system components and enzyme mixtures; storage at –20°C
· Probe Kit (C): Includes target-specific probes, negative and positive control probes, and fluorescent detection probes; storage at –20°C
· Target Retrieval Kit (D): Target retrieval solution; storage at room temperature

Product Advantages
· High Sensitivity: Each target gene is designed with 5–10 probe pairs. Signal amplification via rolling circle amplification (RCA) enhances detection sensitivity and signal-to-noise ratio.
· High Specificity: Dual-ligation probe design ensures highly specific target binding and suppresses amplification of nonspecific hybridization signals.
· Preservation of Tissue Morphology: No region-of-interest selection is required. Whole-slide panoramic detection allows spatial gene expression analysis within complex tissue environments.
· Quantification at Single-Cell Level: The combination of high sensitivity and preserved morphological features enables single-molecule detection at single-cell resolution.
· Simple Operation - No Hybridization Oven Required: Only a constant-temperature incubator, vortex mixer, and mini centrifuge are required. A hybridization oven is not necessary.
· Compatible Sample Types: Fresh frozen tissue sections; FFPE tissue sections; Post-fixed frozen tissue sections; Cultured cell slides

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Background

The SEERNA® FISH RNA Fluorescence In Situ Hybridization Kit is based on signal-amplified single-molecule RNA fluorescence in situ hybridization technology.
First, specially designed paired DNA dual-ligation probes (DLPs) hybridize complementarily to the target RNA molecules. A ligase with RNA-templated DNA ligation capability then catalyzes the first ligation step, joining the DLPs into a semi-circular structure.
Subsequently, using a complementary DNA strand as a template, the DLPs are circularized under the action of a Splint ligase to form a closed DNA circle.
Next, rolling circle amplification (RCA) is carried out by DNA polymerase, generating amplified DNA products and achieving significant signal enhancement.
Finally, fluorescent detection probes labeled with a single fluorophore hybridize to the RCA products, and fluorescence imaging is performed using a fluorescence microscope.