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Cat. No.: FSHKR-100 (for 100T)

Cat. No.: FSHKR-500 (for 500T)

Description

Fluorescence in Situ Hybridization Kit for RNA (FISH Kit for RNA) is a high-sensitivity detection reagent kit used to visualize and determine the cellular localization of target RNA in cells or tissues. It operates on the principle of base complementarity by hybridizing fluorescence-labeled oligonucleotide probes (DNA or RNA) with the target RNA in cells or tissues. This results in the formation of fluorescently labeled nucleic acid probe-target RNA hybrid complexes, which can be observed and analyzed under a fluorescence microscope.

Components

• Proteinase K (20mg/ml)
• Acetylation Solution A
• Acetylation Solution B
• Hybridization Solution
• Yeast RNA (100X)
• DAPI (5mg/ml)
• Wash Buffer (20X)

Reagents that need to be prepared

• Ultrapure Water (DNase/RNase-Free, Sterile) or DEPC Water
• 4% Paraformaldehyde Fixative Solution
• PBS, pH7.3 (DNase, RNase & Protease free, Sterile)
• Hydrochloric Acid
• Deionized Formamide
• Anti-Fluorescence Quenching Mounting Solution
• Poly-L-Lysine (prepare as needed)
• Cover Glasses (prepare as needed)
• Reagents related to frozen or paraffin sections (prepare as needed)
• Target RNA Fluorescent Probes (must be purchased or synthesized independently)

Features

• The FISH Kit for RNA offers a simple and convenient operation. The detection is typically completed within a day. After the fluorescence-labeled nucleic acid probe in the kit forms a hybrid complex with the target RNA, direct fluorescence observation can be conducted.
• The kit exhibits low background and high sensitivity. It provides specific acetylation reagents and additional blocking reagent Yeast RNA (100X) to effectively reduce background and enhance detection sensitivity.
• The kit is applicable to a wide range of samples. It can be used for the localization analysis of target RNA in samples such as tissues fixed with formaldehyde or formalin, paraffin or frozen sections, and cells. The kit is suitable for heterogeneous sample analysis, highly sensitive detection of non-coding RNA (including miRNA and lncRNA) at the single-cell level, tracking detection of RNA from the genome source, and high-throughput single-cell quantitation of target RNA.
• The kit can provide information on the single-cell spatial distribution of target RNA. Gene expression analysis is often performed using techniques such as transcriptome sequencing, qPCR, Northern blotting, and gene microarray chips. Although these methods provide abundant information, they can easily mask the heterogeneity of gene expression at the single-cell level, particularly the spatial distribution of RNA within cells. The FISH Kit for RNA can selectively and highly sensitively reveal the spatial distribution of gene expression at the single-cell level.

Storage

Store at -20°C, with a shelf life of one year. Proteinase K (20mg/ml) and Yeast RNA (100X) should be stored at -20°C. Hybridization Solution and DAPI should be stored at -20°C, protected from light. Acetylation Solution A, Acetylation Solution B, and Wash Buffer (20X) can be stored at 4°C or room temperature.

Note

• It is recommended to use fresh samples for FISH testing whenever possible. Fixed samples that have been stored over time may undergo gradual degradation of RNA, which can affect the effectiveness of FISH detection.
• For routine detection of mammalian cell or tissue samples, it is common to add Yeast RNA to the Hybridization Solution before use to effectively reduce background. However, in cases where Yeast RNA may cause interference, it can be omitted.
• It is advisable to aliquot the Hybridization Solution and store it protected from light.
• Once the pre-hybridization temperature is determined, it is generally recommended to maintain consistent temperatures for subsequent steps, including probe denaturation, hybridization, and washing.
• Each reagent must be completely thawed and mixed well before use.
• Due to the involvement of RNA, it is crucial to strictly follow the guidelines for RNA handling to avoid RNase contamination. The pipette tips, centrifuge tubes, and any equipment or solutions used in the experiment that come into contact with the samples must be RNase-free. Glass or plasticware, instruments, and surfaces in the laboratory should also be treated to be RNase-free whenever possible.
• This product is intended for scientific research purposes by professionals only. It is not to be used for clinical diagnosis or treatment, nor for food or drug purposes. It should not be stored in residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves while handling.

Related: Biotin Fluorescence in Situ Hybridization Kit for RNA (Green)

Only for research and not intended for treatment of humans or animals

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