RNA FISH was performed with the FISH kit according to the manufacturer’s protocol (Ribo Bio Technology, C10910). Cells were seeded in a 24-well plate. When the cell density reached 50–60%, the medium was discarded and 4% paraformaldehyde was added. After washing with PBS, 500 μL of pre-cooled permeate solution was added into cell samples for 3 min. Then permeate solution was washed away with PBS, 200 μL pre-hybridization buffer was added to each well and incubated for 30 min. Subsequently, 100 μL 20 μM lncIAPF, RNA18S and RNU6 FISH Probe Mix were added to each well and incubated overnight. Cells were washed with SSC (Solarbio, S1030) and PBS solution, and stained with DAPI staining solution for 10 min. Finally, the images were observed with a confocal microscope.
S1030
Manufactured by Solarbio
Sourced in China
The S1030 is a laboratory equipment designed for high-throughput protein purification. It features a compact design and automated operation for efficient separation and collection of target proteins from complex samples.
Automatically generated - may contain errors
2 protocols using s1030
1
RNA FISH Protocol for lncRNA Detection
2
Detailed Fluorescence Imaging of lncRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
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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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