Four synthetic RNA oligonucleotides (Integrated DNA Technologies, Coralville, IA, USA), representing the four hsa-miR-21-5p isomiRs were designed (Table 1 ). Each synthetic RNA oligonucleotide bore standard modifications as are seen in the cell: a 5′-phosphate group with a 3′-hydroxyl group. We created four “synthetic cell lines” from these isomiRs, by combining 50 microliters of a 10–14 M starting concentration of the “dominant” isomiR together with 10 microliters of a 10–14 M starting concentration of the other three isomiRs, in H2O. We repeated this process an additional three times to create a synthetic cell line with each of the four isomiRs being “dominant” in turn.
Synthetic rna oligonucleotides
Manufactured by Integrated DNA Technologies
Sourced in United States
Synthetic RNA oligonucleotides are short, single-stranded RNA molecules that are chemically synthesized. They can be used as research tools in various applications, such as gene expression analysis, RNA interference, and RNA-based therapeutics.
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Lab products found in correlation
3 protocols using synthetic rna oligonucleotides
1
Synthetic Cell Lines with miR-21-5p IsomiRs
2
RNA-Protein Binding Assay Protocol
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Gel mobility shift assays were performed as previously described (Williams-Carrier et al. 2008 (link)). Synthetic RNA oligonucleotides (Integrated DNA Technologies) were 5′ end labeled with T4 polynucleotide kinase and [γ-32P]ATP. Binding reactions contained 15 pM RNA, 40 mM Tris–HCl, pH 7.5, 180 mM NaCl, 10% glycerol, 4 mM DTT, 10 U RNAsin, 0.1 mg/mL BSA, 0.5 mg/mL heparin and protein at the indicated concentrations. Binding reactions were incubated for 30 min at 25°C and resolved on 5% polyacrylamide gels in 1× THE (34 mM Tris, 66 mM HEPES, 0.1 mM EDTA, pH 7.5) at 4°C. The data were visualized with a phosphorimager and quantified with ImageQuant. Binding curves were generated with KaleidaGraph software.
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Synthetic isomiR Cell Lines
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