Product description: T4 RNA Ligase 1 is an ATP-dependent ligase, active on a broad range of substrates including RNA, DNA, oligoribonucleotides,oligodeoxynucleotides, as well as numerous nucleotide derivatives. The enzyme catalyzes the formation of a phosphodiester bond between a 5'-phosphoryl-terminated nucleic acid donor to a 3'-hydroxyl-terminated nucleic acid acceptor in a template-independent manner.
Concentration and Size: 10U/μL
Unit Definition: One unit is defined as the amount of enzyme required to ligate 50% of 0.4µg of an equimolar mix of two single stranded 23 base RNA oligonucleotides (one 5'-phosphorylated) in a total volume of 20µL following a 30 minute incubation at 37°C.
Storage Buffer: 10mM Tris-HCl, 0.1mM EDTA, 50mM KCl, 1mM DTT, 50%(v/v)glycerol, pH7.5@25℃.
T4 RNA ligase 1, T4 RNA ligase 2, 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.
Note: T4 RNA ligase 1 requires ATP. (ATP, 100mM Solution, Cat#R1331)
Storage Conditions: Stored at -20±5℃. Do not store in a frost-free freezer. Always avoid freeze-thaw cycles or exposure to frequent temperaturechanges. These fluctuations can greatly alter product stability.
Quality Assurance:
1. Contaminant Assay: Free of Endonuclease, Exonuclease and RNase activities.
2. Endotoxin Assay: Pass.
3. Physical Purity: >90%.
