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Ribonuclease R (RNase R)
Product Details
ShowRibonuclease R from E. Coli includes the following components based on size:
250 units:
12.5 μL of the RNase R enzyme and 250 μL of the optimised 10X Reaction Buffer containing 0.2 M Tris-HCl (pH 8.0), 1 M KCl, and 1 mM MgCl2.
2500 units:
125 μL of the RNase R enzyme and 2.5 mL of the optimised 10X Reaction Buffer containing 0.2 M Tris-HCl (pH 8.0), 1 M KCl, and 1 mM MgCl2.
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Ribonuclease R (RNase R)
Ribonuclease R (RNase R), 250 Units
Ribonuclease R (RNase R) from E. coli is a magnesium-dependent 3ʹ to 5ʹ exoribonuclease available for circular RNA cleanup, alternative splicing research, and gene expression studies.
Key features
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Efficacy and quality assured under ISO 13485 certification
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Available in bulk and for custom configurations
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Effectively degrades linear RNA, Y-structure RNAs, but not lariat loops or circular RNAs.
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Digests linear RNAs to enrich for circular RNAs used for protein production or intronic cDNA library construction.
Product information
Ribonuclease R (RNase R) from E. coli is a magnesium-dependent 3′→ 5′ exoribonuclease that digests linear RNAs. RNase R does not digest lariat or circular RNA structures.1,2 Most cellular RNAs will be digested completely, except for tRNAs, 5S RNA, and intron lariats.
Lariats are produced during pre-mRNA splicing of intron regions (Figure1) and the 3´ tails will be trimmed by RNase R to the branch point nucleotide, where there is a 2´,5´-phosphodiester linkage.
RNase R Applications:
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Removal of precursor linear RNA after circularisation of RNA for enhanced protein production
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Alternative splicing studies
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Gene expression studies
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Intron cDNA production
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Intronic screening of cDNA libraries
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Isolation of splicing intermediates and lariats
Figure 1. Schematic overview showing processing of intron lariats by RNase R.
RNase R is also used in the identification of circular RNA for producing therapeutics and identifying certain viral mechanisms.3,4 CircRNA has been referred to as the “new generation of mRNA therapy” and can be confirmed and validated for further analysis and sequencing using RNase R treatment.3,4 RNase R treatment is also used in identifying RNase R resistant circRNA and to enrich circRNA by selective degradation of linear RNAs.5,6
References
- Suzuki, H. et al. (2006) Nucleic Acids Res. 34, 63.
- Vincent, H.A. and Deutscher, M. P. (2006) J. Biol. Chem. 281, 29769.
- Chen, C., et al. (2022). bioRxiv, 2022-05.
- Chasseur, A. S., et al. (2022). J. Virol, 96(9), e00321-22.
- Chen, R., Wang, S.K., Belk, J.A. et al. (2022). Nat Biotechnol 41, 262–272.
- Breuer J, Barth P, Noe Y, et al. (2022). Mol. Ther. Nucleic Acids, 28, 623-635.
Note: RNase R requires low (0.1-1.0 mM) magnesium concentrations for activity. Low EDTA concentrations in substrate RNA solutions can negatively affect RNase R activity. Additional MgCl2 up to 1 mM final concentration can be used to compensate for EDTA in the substrate. Optimal activity is at 37 °C
Concentration: 20 U/ μL
Unit Definition: One unit of RNase R converts 1 µg of poly(A) into acid-soluble nucleotides in 10 minutes at 37 °C under standard assay conditions.
Storage Buffer: RNase R is supplied in a 50% glycerol solution containing 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton® X-100.
RNase R 10X Reaction Buffer: 0.2 M Tris-HCl (pH 8.0), 1 M KCl, and 1 mM MgCl2.
SDS
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10X RNase R Reaction Buffer English EU
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10X RNase R Reaction Buffer English US
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RNase R English EU
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RNase R English US
Manuals and user guides
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