Product description: Ribonuclease R (RNase R) from E. coli, is a magnesium-dependent 3' to 5' exoribonuclease that digests essentially all linear RNAs but does not digest lariat or circular RNA structures, or double-stranded RNA with 3'-overhangs shorter than 7 nucleotides. RNase R digests RNA from the 3' to 5' direction, releasing 5'-monophosphate nucleotides to produce dinucleotide and trinucleotide end products.
Resource: Recombinant E.coli
Concentration and Size: 20U/μL
Unit Definition: One unit converts 1μg of poly-r(A) into acid-soluble nucleotides in 10 minutes at 37℃.
Storage Buffer: 50mM Tris-HCl, 0.1mM EDTA, 100mM NaCl, 1mM DTT, 0.1%(v/v)Triton X-100, 50%(v/v) glycerol, pH7.5@25℃.
T4 RNA ligase1, T4 RNA ligase2, RNaseR on enzymatic synthesis of circular RNA:
IVT reaction to obtain linear RNA: It is worth noting that for this step of IVT reaction, a certain proportion of GMP needs to be doped to obtain GMP-containing linear RNA;
RNA cyclisation: circular RNA is produced by joining the two ends within the RNA molecule by T4 RNA ligase 1 (T4 Rnl 1) or T4 RNA ligase 2 (T4 Rnl 2) catalysing the joining of 5'-phosphate and 3'-hydroxyl groups at the RNA terminus;
Removal of linear RNA: RNaseR is able to remove all linear RNA molecules from the reaction solution, resulting in linear RNA-free circular RNA.
Storage Conditions: Stored at -20±5°C. Do not store in a frost-free freezer. Always avoid freeze-thaw cycles or exposure to frequent temperature changes. These fluctuations can greatly alter product stability.
Quality Assurance:
1. Functional Assay: Passes the internal function testing.
2. Contaminant Assay: Free of detectable Endonuclease and Exonuclease.
3. Endotoxin Assay: Pass.
4. Physical Purity: >95% by SDS-PAGE with Coomassie® blue staining.
