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.

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• Effectively degrades linear RNA, Y-structure RNAs, but not lariat loops or circular RNAs.
• Digests linear RNAs to enrich for circular RNAs used for protein production or intronic cDNA library construction.

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