SEERNA® CISH RNA Chromogenic In Situ Hybridization Kit

The SEERNA® CISH RNA Chromogenic In Situ Hybridization Kit is developed based on single-molecule chromogenic in situ hybridization (smCISH) technology. This technique is built upon DNA padlock probe design and rolling circle amplification (RCA), enabling highly specific in situ localization and validation of single-target gene expression, including mRNA and selected lncRNAs, within tissue samples.

The kit offers high detection efficiency, high specificity, and a shortened experimental workflow.

It is particularly suitable for samples with high autofluorescence background and no requirement for multiplex co-detection. For example, it is ideal for validating RNA expression of a specific target or biomarker in clinical FFPE samples. Additionally, it can be applied to tissue detection of exogenous infectious viruses such as HIV, HCV, and HPV.

Features
· Delivers high detection efficiency, strong specificity, and reduced experimental time.
· Applicable in neuroscience, oncology, developmental biology, and drug development. The system is user-friendly and requires lower hardware and software specifications.
· Suitable for various tissue types (human and mouse tissues from multiple sources) and multiple sample preparation methods, enabling coverage of conventional sample types.
· Particularly suitable for samples with high autofluorescence background and no need for multiplex detection.

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Background

This technology is developed based on padlock probes and does not require reverse transcription. It enables highly sensitive and highly specific detection of target RNA molecules in fixed cells or tissue samples.
A padlock probe is a linear single-stranded DNA molecule composed of two target-recognition arms at its 5′ and 3′ ends, along with a non-target-dependent central sequence. Only when the probe hybridizes perfectly and complementarily to the target RNA can it be ligated into a circular molecule by DNA ligase. The circularized product then serves as a template for rolling circle amplification (RCA), generating a large number of amplification products.
Subsequently, the amplified products hybridize with an HRP-labeled detection probe. In the presence of the substrate DAB, HRP catalyzes the formation of a brown precipitate, which can be visualized under a bright-field microscope.
By quantifying the amount of brown precipitate within cells, the expression level of the target RNA can be evaluated.