RNA-FISH probes, including circANKRD42, U6, and 18S FISH probes, were synthesized by Ribo Bio Technology (Guangzhou, China). MRC-5 cells were inoculated on a circular slide, fixed with 4% paraformaldehyde, washed with PBS, and punched with 0.5% Triton X-100. After incubation at 4°C for 3 min, pre-hybridization solution was dropped. Then, the denatured probe mix was added to the culture well and placed in hybridization oven at 37°C in the dark overnight. The next day, the hybridization probe solution was aspirated at 42°C and the slides were rinsed sequentially with 2× saline sodium citrate (SSC) and 1× SSC and washed three times for 5 min each. We added 150 μL DAPI solution, stained for 6 min at room temperature, and washed with PBS. Fluorescence was observed by a laser confocal microscope. The FISH probe sequence of circANKRD42 and miR-324-5p was hsa_circANKRD42 (CY3 labeled): 5ʹ-gaagacactctttgccacatcactgggt-3ʹ and hsa_miR-324-5p (fluorescein isothiocyanate [FITC] labeled): 5ʹ-accaatgccctaggggatgcg-3ʹ.
18s fish probes
Manufactured by RiboBio
Sourced in China
18S FISH probes are fluorescent in situ hybridization (FISH) probes that target the 18S ribosomal RNA (rRNA) gene. The 18S rRNA is a component of the small subunit of eukaryotic ribosomes and is widely used as a marker for identifying and quantifying specific cell types or microorganisms in various sample types.
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Lab products found in correlation
4 protocols using 18s fish probes
1
RNA-FISH Analysis of circANKRD42 and miR-324-5p
2
FISH Assay for Gene Expression
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OIP5-AS1, U6, and 18S FISH probes were purchased from RiboBio (Guangzhou, China), and the FISH kit was used to perform the FISH experiment (RiboBio). Cells fixed with 4% polyoxymethylene were incubated with 0.5% Triton X-100 solution at 4 °C for 5 min. Cells were washed thrice with PBS before being treated with pre-hybridization buffer at 37 °C for 30 min. After removing the pre-hybridization buffer, a 20 µM probe mix was added and incubated overnight at 37 °C. Cells were stained with DAPI for 10 min, sealed, and observed under a confocal microscope (Leica, Solms, Germany).
3
Fluorescent in situ Hybridization in Fish
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The loc339803, U6 and 18S fish probes were designed and synthesized by Guangzhou Ribobio Co, Ltd. (Guangzhou, China). The fish excrement was carried out by using the fish kit according to the manufacturer's instructions. The images were acquired through confocal microscope (Leica).
4
Detailed Fluorescence Imaging of lncRNA
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lncITPF, U6, and 18S FISH probes were synthesized by Guangzhou RiboBio Co., Ltd. The experiment was performed with the Ribo lncRNA FISH Probe Mix according to the manufacturer’s protocol (RiboBio, C10910, Guangzhou, China). Cells were inoculated on a circular slide. When the cell density reached 50–60%, 4% paraformaldehyde (Meilunbio, MA0192, Dalian, China) was added. Then, cell samples were washed with 1 × PBS (Sparkjade, CR0013, Jinan, China) and punched with 0.3% TritonX-100 (Sinopharm, 30188928, Shanghai, China) at 4 °C for 3 min. Permeate solution was washed away with 1 × PBS and 200 μL pre-hybridization solution was added for 30 min. Subsequently, lncITPF, U6, and 18S FISH Probe Mix were added to the cell samples in a hybridization oven at 37 °C overnight. On the second day, the hybridization probe solution was aspirated at 42 °C and washed with SSC (Solarbio, S1030, Beijing, China). Then DAPI (Sigma, D9542, St. Louis, MO, USA) solution was added for 6 min. Finally, fluorescence was observed using a laser confocal microscope.
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