Ribonuclease R (RNase R) from E. coli, is a magnesium-dependent 3′→5′ exoribonuclease that can digest all linear RNAs but does not digest lariat or circular RNA structures. Most cellular RNAs will be digested completely by RNase R, with the exception of tRNAs, 5S RNA and intron lariats. RNase R is used for enrichment of circRNAs in biological samples, identification of intronic lariat sequences, identification of exonic circRNAs, studying alternative splicing. RNase R must be used in combination with the 10× RNase R Reaction Buffer, GMP-grade (Cat. No. 14616ES) Features Guaranteed efficacy and quality with ISO 13485 certification. Available in bulk or tailored configurations. Effectively breaks down linear RNA, while preserving lariat loops and circular RNAs. Selective digestion of linear RNAs enhances the isolation of circular RNAs for protein production or intronic cDNA library construction. Offering GMP-Grade RNAse R Specificntions Source A recombinant E. coli strain carrying the RNAse R gene from E. coli Optimum Temperature 37℃ Storage Buffer 50 mM Tris-HCl,100 mM NaCl, 0.1 mM EDTA, 0.1% Triton® X-100, 1 mM DTT, 50% glycerol，pH7.5 at 25℃ Unit Definition One unit converts 1 μg of poly-r(A) into acid-soluble nucleotides in 10 minutes at 37°C in 20 mM Tris-HCl (pH 8.0), 100 mM KCl and 0.1 mM MgCl2. Application RNase R serves as a fundamental component in circular RNA synthesis (circRNA). CircRNA holds promise for enhancing pharmaceutical stability and biostability. With their covalently closed structures, circRNAs withstand RNA exonucleases. [1,2] RNase R finds widespread application in various studies, including alternative splicing, intron cDNA production, and the isolation of splicing intermediates and lariats. Components Components No. Name 14615ES25 14615ES72 14615ES80 14615 RNase R (GMP-grade, 20 U/μL) 25 μL 250 μL 1 mL Storage This product should be stored at -25~-15℃ for 2 years. Instructions Prepare the following reaction system in a sterile microcentrifuge tube component Volume 10×RNase R Reaction Buffer 2 μL RNA sample 1 μg RNase R (20 U/μL) 2-4 U RNase-free ddH2O Up to 20 μL Incubate at 37 ℃ for 10 - 30 minutes. Figures Figure 1: Total mRNA (C) can be digested by Yeasen RNAse R. Total mRNA input: 500ng. Figure 2: CircRNA can not be digested by Yeasen RNAse R. CircRNA input: 500ng, RNAse R: 4U. Figure 3: Performence of Yeasen RNAse R (1U to 8U ) digestion on 1 μg of linear mRNA (C). Notes The activity of RNase R requires 0.1-1.0 mM Mg2+; With the increase of RNA, the reaction time can be extended and the enzyme amount can be increased appropriately; The EDTA content in RNA samples may affect the activity of RNase R. Incubate at 70 ℃ for 10 minutes can inactivate the enzyme. Reference Qu L, Yi Z, Shen Y, et al. Circular RNA vaccines against SARS-CoV-2 and emerging variants. Cell. 2022;185(10):1728-1744.e16. doi:10.1016/j.cell.2022.03.044 Chen L, Wang C, Sun H, et al. The bioinformatics toolbox for circRNA discovery and analysis. Brief Bioinform. 2021;22(2):1706-1728. doi:10.1093/bib/bbaa001 Documents Safety Data Sheet 14615_MSDS_HB260713 Manual RNAse R GMP-Grade manual
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