T4 RNA Ligase 2 catalyzes phosphodiester bond formation between a 5′ phosphate and 3′ hydroxyl of RNA. The preferred substrate is nicked double-stranded RNA but single-stranded RNA can also serve as a substrate. Ligation of single-stranded RNA substrates generates either intramolecular or intermolecular products. Besides nicked double-stranded RNA substrates, other nicked nucleic acids hybrids can be sealed. The strand containing the 5′ phosphate can either be DNA or RNA. The non-ligated strand of the duplex can be either RNA or DNA. T4 RNA ligase 2 requires ATP for activity unless the substrate is preadenylated on the 5′ end. A truncated version of T4 RNA ligase 2 is a better enzyme for preadenylated substrates because it generates less side-reaction ligation products than the full length enzyme.
Source of ProteinPurified from a strain of E. coli that expresses the recombinant T4 RNA Ligase 2 gene.
Supplied in10 mM Tris-HCl100 mM NaCl0.1 mM DTT0.1 mM EDTA50% glycerolpH 7.5 @ 25°C
Supplied With:B6030 (10X Ligation Buffer)
10X Ligation Buffer (B6030)500 mM Tris-HCl100 mM MgCl250 mM DTT10 mM ATPpH 7.6 @ 25°C
Unit DefinitionOne unit is defined as the amount of enzyme required to ligate 50% of 0.4 µg of an equimolar mix of a single-stranded 5′ FAM-labeled 17-mer RNA to the 5′ phosphorylated end of a 18-mer DNA when both strands are annealed to a complementary 35-mer DNA strand in 20 µL at 37°C for 30 minutes.
